Fluorinated tetrahydronaphthyridinyl nonanoic acid derivatives and uses thereof

ABSTRACT

The present invention relates to fluorinated compounds of formula I and methods of synthesizing these compounds. The present invention also relates to pharmaceutical compositions containing the fluorinated compounds of the invention, and methods of treating fibrosis, macular degeneration, diabetic retinopathy (DR), macular edema, diabetic macular edema (DME), and macular edema following retinal vein occlusion (RVO), by administering these compounds and pharmaceutical compositions to subjects in need thereof.

RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.16/373,780, filed Apr. 3, 2019, which is a continuation of U.S.application Ser. No. 15/700,351, filed Sep. 11, 2017 (now U.S. Pat. No.10,301,307), which is a continuation of U.S. application Ser. No.15/047,717, filed Feb. 19, 2016 (now U.S. Pat. No. 9,790,222), whichclaims the benefit of and priority to U.S. provisional application No.62/118,303, filed Feb. 19, 2015, the entire contents of each of whichare incorporated herein by reference in their entireties.

BACKGROUND OF THE INVENTION

Fibrosis is characterized by excessive accumulation of collagen in theextracellular matrix of the involved tissue. It is a long-standing andchallenging clinical problem for which no effective treatment iscurrently available. The production of collagen is a highly regulatedphysiological process, the disturbance of which may lead to thedevelopment of tissue fibrosis. The formation of fibrous tissue is partof the normal beneficial process of healing after injury. In some cases,however, an abnormal accumulation of fibrous material can severelyinterfere with the normal function of the affected tissue or even causethe complete loss of function of the affected organ.

A variety of compounds have been identified as anti-fibrosis agents viadifferent mechanisms of action, including the suppression of collagenexpression. For example, pantethine(D-bis-(N-pantothenyl-β-aminoethyl)-disulfide) has been reported to beeffective for the inhibition of hepatic fibrosis (U.S. Pat. No.4,937,266). Also, a hydrazine derivative, benzoic hydrazide, has beenshown to be a powerful antifibrotic agent (U.S. Pat. Nos. 5,374,660 and5,571,846). In addition, angiotensin inhibitors are used in combinationwith nitric oxide stimulators to inhibit the progression of fibrosis(U.S. Pat. Nos. 5,645,839 and 6,139,847). Further, A₁ adenosine receptorantagonists and/or P_(2x) purinoceptor antagonists are described fortreating or preventing fibrosis and sclerosis (U.S. Pat. No. 6,117,445).More recently, somatostatin agonists, hepatocyte growth factors (HGFs),chymase inhibitors, and antagonists of IL-13 have been reported toeffectively inhibit fibrosis (U.S. Pat. Nos. 6,268,342, 6,303,126,6,500,835, and 6,664,227).

Age-related macular degeneration (AMD) is the leading cause of blindnessin people over 55; and diabetic retinopathy (DR) is the leading cause inpeople under 55 (Klein, 1994; Williams, 2004). Both diseases arecharacterized by new blood vessel growth (Freund, 1993; Speicher, 2003;Zarbin, 2004). Macular edema and Diabetic macular edema (DME) occur whenfluid and protein deposits collect on or under the macula caused byleaking macular capillaries. Thrombosis of central retinal vein (CRV)and its branches is the second most prevalent vascular pathology afterDR, and results in abrupt decrease in visual acuity and is accompaniedby macular edema. Thus, anti-angiogenesis treatments are useful incombating all these conditions.

Integrins are heterodimeric transmembrane proteins through which cellsattach and communicate with extracellular matrices and other cells. αvintegrins are key receptors involved in mediating cell migration andangiogenesis. αv integrins have been shown to be involved in a number ofdiseases and conditions including ocular angiogenesis and fibrosis oforgans. Expression of αv integrins is upregulated in various diseases orconditions, such as AMD and DR, and in mouse model of oxygen-inducedretinopathy (OIR) or retinopathy of prematurity (ROP) model (Takagi,2002). Also, αvβ3 is expressed in new vessels after photocoagulation,but not in normal choroidal vessels, in the laser-induced choroidalneovascularization model for AMD (Kamizuru, 2001). Administration of αvintegrins antagonists, such as a cyclic RGD peptide, has been shown toinhibit retinal and choroidal neovascularization (Friedlander, 1996;Chavakis, 2002; Luna, 1996; Riecke, 2001; Yasukawa, 2004). Angiogenesisinhibitors targeting vascular endothelial growth factor (VEGF), othergrowth factors (e.g., fibroblast growth factor (FGF), platelet-derivedgrowth factor (PDGF)), chemokines (e.g., IL8, SDF1, G-CSF), receptors(e.g., CXCR1, FGF-R, PlGFR, PDGFR, Tie-receptors), intracellularmediators (e.g., c-kit kinase, PI3 kinase, PKC), and extracellularmediators (e.g., integrins, cadherins), as well as inhibitors ofpro-angiogenic targets (e.g., phosphoinositide 3 kinase), have beeninvestigated for the treatment of AMD and DR. However, application ofthese drugs is limited.

Thus, there continues to be a need for compounds, compositions, andmethods for treating fibrosis, AMD, DR, DME, and macular edema followingretinal vein occlusion, that are safe, effective, and convenientlyadministered. The present invention addresses the need.

SUMMARY OF THE INVENTION

The present invention provides a compound of formula I:

or a pharmaceutically acceptable salt or solvate thereof, wherein thecompound of formula I is defined in detail herein below.

The present invention also provides a pharmaceutical compositioncomprising a compound of formula I or a pharmaceutically acceptable saltor solvate thereof, and a pharmaceutically acceptable carrier orexcipient.

The present invention also provides a method of treating or preventing afibrosis, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of formula Ia:

or a pharmaceutically acceptable salt or solvate thereof or atherapeutically effective amount of a pharmaceutical composition of theinvention, wherein the compound of formula Ia is defined in detailherein below. In one aspect, the invention provides treating a fibrosis.In one aspect, the invention provides preventing a fibrosis.

The present invention also provides the use of a compound of formula Iaor a pharmaceutically acceptable salt or solvate thereof in themanufacture of a medicament for the treatment or prevention of afibrosis in a subject. The present invention also provides the use of acompound of formula Ia or a pharmaceutically acceptable salt or solvatethereof in treating or preventing a fibrosis in a subject.

The present invention also provides a method of treating or preventing adisease or condition in a subject, comprising administering to a subjectin need thereof a therapeutically effective amount of a compound offormula I or a pharmaceutically acceptable salt or solvate thereof or atherapeutically effective amount of a pharmaceutical compositioncomprising a compound of formula I or a pharmaceutically acceptable saltor solvate thereof. In one aspect, the invention provides treating adisease or condition. In one aspect, the invention provides preventing adisease or condition.

The present invention provides a method of treating or preventing adisease or condition mediated by an αv integrin in a subject, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula I or a pharmaceutically acceptable saltor solvate thereof or a therapeutically effective amount of apharmaceutical composition comprising a compound of formula I or apharmaceutically acceptable salt or solvate thereof. In one aspect, thedisease or condition is a disease or condition in which angiogenesis isinvolved. In a further aspect, the disease or condition is a disease orcondition in which ocular angiogenesis is involved.

The present invention also provides a method of treating or preventingan αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin-mediated disease or conditionin a subject, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of formula I or apharmaceutically acceptable salt or solvate thereof or a therapeuticallyeffective amount of a pharmaceutical composition comprising a compoundof formula I or a pharmaceutically acceptable salt or solvate thereof.In one aspect, the disease or condition is a disease or condition inwhich ocular angiogenesis is involved. In one aspect, the disease orcondition is macular degeneration. In one aspect, the disease orcondition is age-related macular degeneration (AMD). In one aspect, thedisease or condition is diabetic retinopathy (DR). In one aspect, thedisease or condition is diabetic macular edema (DME). In one aspect, thedisease or condition is macular edema following retinal vein occlusion(RVO). In one aspect, the condition is fibrosis of the liver, kidney,intestine, lung, and heart. In one aspect, the disease is a renaldisease, a respiratory disease, a gastrointestinal disease, acardiovascular disease, a bone and articular disease, a skin disease, anobstetric disease, or a urologic disease.

The present invention provides the use of a compound of formula I or apharmaceutically acceptable salt or solvate thereof in the manufactureof a medicament for the treatment or prevention of a disease orcondition in a subject. The present invention provides the use of acompound of formula I or a pharmaceutically acceptable salt or solvatethereof in treating or preventing a disease or condition in a subject.

The present invention provides the use of a compound of formula I or apharmaceutically acceptable salt or solvate thereof in the manufactureof a medicament for the treatment or prevention of a disease orcondition mediated by an αv integrin in a subject. The present inventionprovides the use of a compound of formula I or a pharmaceuticallyacceptable salt or solvate thereof in treating or preventing a diseaseor condition mediated by an αv integrin in a subject. In one aspect, thedisease or condition is a disease or condition in which angiogenesis isinvolved. In a further aspect, the disease or condition is a disease orcondition in which ocular angiogenesis is involved.

The present invention also provides the use of a compound of formula Ior a pharmaceutically acceptable salt or solvate thereof in themanufacture of a medicament for the treatment or prevention of an αvβ3,αvβ5, αvβ6 and/or αvβ8 integrin-mediated disease or condition in asubject. The present invention provides the use of a compound of formulaI or a pharmaceutically acceptable salt or solvate thereof in treatingor preventing of an αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin-mediateddisease or condition in a subject. In one aspect, the disease orcondition is a disease or condition in which ocular angiogenesis isinvolved. In one aspect, the disease or condition is maculardegeneration. In one aspect, the disease or condition is age-relatedmacular degeneration (AMD). In one aspect, the disease or condition isdiabetic retinopathy (DR). In one aspect, the disease or condition isdiabetic macular edema (DME). In one aspect, the disease or condition ismacular edema following retinal vein occlusion (RVO). In one aspect, thecondition is fibrosis of the liver, kidney, intestine, lung, and heart.In one aspect, the disease is a renal disease, a respiratory disease, agastrointestinal disease, a cardiovascular disease, a bone and articulardisease, a skin disease, an obstetric disease, or a urologic disease.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. In the case of conflict, thepresent specification, including definitions, will control. In thespecification, the singular forms also include the plural unless thecontext clearly dictates otherwise. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference. The references cited herein are not admitted to be prior artto the claimed invention. In addition, the materials, methods, andexamples are illustrative only and are not intended to be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows lung stiffness as measured by the pressure volume curves inmice when dosed with 10 mg/kg, 30 mg/kg, and 100 mg/kg of Compound A15s,pirfenidone, saline, or vehicle.

FIG. 2 shows lung stiffness as measured by the pressure volume curves inmice when dosed with 10 mg/kg, 30 mg/kg, and 100 mg/kg of Compound A21,pirfenidone, saline, or vehicle.

DETAILED DESCRIPTION OF THE INVENTION

Compounds of the Invention

The present invention relates to novel fluorinated compounds of formulaI:

or a pharmaceutically acceptable salt or solvate thereof, wherein:

Z is

Q is

X is CR₄ or N;

Y is CR₄ or N;

R₁ is H, F, Cl, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms;

R₂ and R₃ are each independently H, F, CH₂F, CHF₂, or CF₃, provided thatone of R₂ and R₃ is not H;

each R₄ is independently H, CH₂F, CHF₂, or CF₃; and

R₅₁ and R₅₂ are each independently H, F, or Cl;

provided that the compound of formula I contains at least one fluorineatom, and provided that when Z is

R₁ is not H, F, or Cl, and R₅₁ and R₅₂ are each H, then at least one ofX and Y is CR₄, and R₄ is CH₂F, CHF₂, or CF₃.

The compounds of the present invention contain at least one fluorineatom. In one aspect, the compounds of the present invention contain atleast one fluorine atom in the R₁ substituent. In another aspect, thecompounds of the present invention contain at least one fluorine atom inthe R₂ or R₃ substituent. In another aspect, the compounds of thepresent invention contain at least one fluorine atom in the R₄substituent.

In one aspect, Z is

In another aspect, Z is

In another aspect, Z is

In one aspect, Q is

In one aspect, X is N and Y is CR₄. In another aspect, X and Y are eachCR₄. In another aspect, X and Y are each N.

In one aspect, at least one R₄ is H. In one aspect, at least one R₄ isCH₂F, CHF₂, or CF₃. In a further aspect, at least one R₄ is CF₃.

In one aspect, R₁ is H. In another aspect, R₁ is F, Cl, C₁-C₄ alkylsubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms, or C₁-C₆alkoxy substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms. In afurther aspect, R₁ is F or Cl. In another aspect, R₁ is C₁-C₄ alkylsubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms, or C₁-C₆alkoxy substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms.

In a further aspect, R₁ is straight chain C₁-C₄ or branched C₃-C₄ alkyl,and is substituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms. Ina further aspect, R₁ is methyl, ethyl, propyl, or butyl, and issubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms. In afurther aspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms.In a further aspect, R₁ is CF₃.

In another further aspect, R₁ is straight chain C₁-C₆ or branched C₃-C₆alkoxy, and is substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorineatoms. In a further aspect, R₁ is methoxy, ethoxy, propoxy, or butoxy,and is substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms. In afurther aspect, R₁ is methoxy substituted with 0, 1, 2, or 3 fluorineatoms. In a further aspect, R₁ is OCH₃, OCH₂F, OCHF₂, or OCF₃. In afurther aspect, R₁ is OCHF₂ or OCF₃.

In one aspect, R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ andR₅₂ is H, and the other is F or Cl. In a further aspect, one of R₅₁ andR₅₂ is H, and the other is F. In another aspect, R₅₁ and R₅₂ are each For Cl.

In another aspect, Q is

In one aspect, R₂ is F. In a further aspect, R₂ is F and R₃ is H. Inanother aspect, R₂ is CH₂F, CHF₂, or CF₃.

In one aspect, R₃ is F. In a further aspect, R₃ is F and R₂ is H. Inanother aspect, R₃ is CH₂F, CHF₂, or CF₃. In a further aspect, R₃ isCF₃. In a further aspect, R₃ is CF₃ and R₂ is H.

In one aspect, R₂ and R₃ are each F.

Any of the substituent groups illustrated above for any of X, Y, Z, Q,R₁, R₂, R₃, R₄, R₅₁, and R₅₂ can be combined with any of the substituentgroups illustrated above for the remaining of X, Y, Z, Q, R₁, R₂, R₃,R₄, R₅₁, and R₅₂.

In one aspect, Q is

X is N or CH; Y is CR₄; R₄ is CH₂F, CHF₂, or CF₃; and R₁ is F or Cl. Ina further aspect, R₄ is CF₃; and R₁ is F or Cl. In a further aspect, R₁is F. In another further aspect, R₁ is Cl.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotherfurther aspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atomsor methoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In another furtheraspect, R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotheraspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms ormethoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In a further aspect,R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotheraspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms ormethoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In a further aspect,R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

In one aspect, a compound of present invention is of formula II:

or a pharmaceutically acceptable salt or solvate thereof, wherein eachof the variables is as defined above. Compounds of the present inventioninclude compounds of formula II, wherein the variables and combinationsthereof are illustrated in the various aspects of formula I above.

In one aspect, a compound of present invention is of formula Ma or Mb:

or a pharmaceutically acceptable salt or solvate thereof, wherein Z′ is

and each of the other variables is as defined above. Compounds of thepresent invention include compounds of formula Ma or Mb, wherein thevariables and combinations thereof are illustrated in the variousaspects of formula I above.

In one aspect, Q is

In one aspect, X is N and Y is CR₄. In another aspect, X and Y are eachCR₄. In another aspect, X and Y are each N.

In one aspect, at least one R₄ is H. In one aspect, at least one R₄ isCH₂F, CHF₂, or CF₃. In a further aspect, at least one R₄ is CF₃.

In one aspect, a compound of present invention is of formula IVa or IVb:

or a pharmaceutically acceptable salt or solvate thereof, wherein R₄′ isCH₂F, CHF₂, or CF₃, and each of the other variables are as definedabove. Compounds of the present invention include compounds of formulaIVa or IVb, wherein the variables and combinations thereof areillustrated in the various aspects of formula I above.

In one aspect, R₁ is H, F, or Cl. In a further aspect, R₁ is F or Cl. R₁is Cl.

In one aspect, R₄′ is CF₃.

Representative compounds of the present invention include the compoundslisted in Table 1.

TABLE 1 Cmpd # Chemical Structure A15

A15s

A16

A16s

A17

A17s

A18

A18s

A19

A19s

A20

A20s

A21

A21s

A21-1 enan- tiomer 1

A21-2 enan- tiomer 2

A22

A22s

A23

A23s

A24

A24s

A25

A25s

A26

A26s

A27

A27s

A28

A28s

A29

A29s

A30

A30s

The present invention also relates to use of a compound of formula Ia:

or a pharmaceutically acceptable salt or solvate thereof, for treatingor preventing fibrosis, wherein:

Z is

Q is

X is CR₄ or N;

Y is CR₄ or N;

R₁ is H, F, Cl, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms;

R₂ and R₃ are each independently H, F, CH₂F, CHF₂, or CF₃, provided thatone of R₂ and R₃ is not H;

each R₄ is independently H, CH₂F, CHF₂, or CF₃; and

R₅₁ and R₅₂ are each independently H, F, or Cl;

provided that the compound of formula Ia contains at least one fluorineatom.

The compounds of the present invention contain at least one fluorineatom. In one aspect, the compounds of the present invention contain atleast one fluorine atom in the R₁ substituent. In another aspect, thecompounds of the present invention contain at least one fluorine atom inthe R₂ or R₃ substituent. In another aspect, the compounds of thepresent invention contain at least one fluorine atom in the R₄substituent.

In one aspect, Z is

In another aspect, Z is

In another aspect, Z is

In one aspect, Q is

In one aspect, X is N and Y is CR₄. In another aspect, X and Y are eachCR₄. In another aspect, X and Y are each N.

In one aspect, at least one R₄ is H. In one aspect, at least one R₄ isCH₂F, CHF₂, or CF₃. In a further aspect, at least one R₄ is CF₃.

In one aspect, R₁ is H. In another aspect, R₁ is F, Cl, C₁-C₄ alkylsubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms, or C₁-C₆alkoxy substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms. In afurther aspect, R₁ is F or Cl. In another aspect, R₁ is C₁-C₄ alkylsubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms, or C₁-C₆alkoxy substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms.

In a further aspect, R₁ is straight chain C₁-C₄ or branched C₃-C₄ alkyl,and is substituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms. Ina further aspect, R₁ is methyl, ethyl, propyl, or butyl, and issubstituted with 1, 2, 3, 4, 5, 6, 7, 8, or 9 fluorine atoms. In afurther aspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms.In a further aspect, R₁ is CF₃.

In another further aspect, R₁ is straight chain C₁-C₆ or branched C₃-C₆alkoxy, and is substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorineatoms. In a further aspect, R₁ is methoxy, ethoxy, propoxy, or butoxy,and is substituted with 0, 1, 2, 3, 4, 5, 6, or 7 fluorine atoms. In afurther aspect, R₁ is methoxy substituted with 0, 1, 2, or 3 fluorineatoms. In a further aspect, R₁ is OCH₃, OCH₂F, OCHF₂, or OCF₃. In afurther aspect, R₁ is OCHF₂ or OCF₃.

In one aspect, R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ andR₅₂ is H, and the other is F or Cl. In a further aspect, one of R₅₁ andR₅₂ is H, and the other is F. In another aspect, R₅₁ and R₅₂ are each For Cl.

In another aspect, Q is

In one aspect, R₂ is F. In a further aspect, R₂ is F and R₃ is H. Inanother aspect, R₂ is CH₂F, CHF₂, or CF₃.

In one aspect, R₃ is F. In a further aspect, R₃ is F and R₂ is H. Inanother aspect, R₃ is CH₂F, CHF₂, or CF₃. In a further aspect, R₃ isCF₃. In a further aspect, R₃ is CF₃ and R₂ is H.

In one aspect, R₂ and R₃ are each F.

Any of the substituent groups illustrated above for any of X, Y, Z, Q,R₁, R₂, R₃, R₄, R₅₁, and R₅₂ can be combined with any of the substituentgroups illustrated above for the remaining of X, Y, Z, Q, R₁, R₂, R₃,R₄, R₅₁, and R₅₂.

In one aspect, Q is

X is N or CH; Y is CR₄; R₄ is CH₂F, CHF₂, or CF₃; and R₁ is F or Cl. Ina further aspect, R₄ is CF₃; and R₁ is F or Cl. In a further aspect, R₁is F. In another further aspect, R₁ is Cl.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotherfurther aspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atomsor methoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In another furtheraspect, R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotheraspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms ormethoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In a further aspect,R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In anotheraspect, R₁ is methyl substituted with 1, 2, or 3 fluorine atoms ormethoxy substituted with 0, 1, 2, or 3 fluorine atoms. In a furtheraspect, R₁ is OCHF₂ or OCF₃; X is N; and Y is CH. In a further aspect,R₁ is CF₃; X is N; and Y is N.

In one aspect, Z is

Q is

and R₁ is Cl, F, C₁-C₄ alkyl substituted with 1, 2, 3, 4, 5, 6, 7, 8, or9 fluorine atoms, or C₁-C₆ alkoxy substituted with 0, 1, 2, 3, 4, 5, 6,or 7 fluorine atoms. In a further aspect, R₁ is Cl or F. In a furtheraspect, R₄ is CH₂F, CHF₂, or CF₃. In a further aspect, X is CH; Y isCR₄; R₁ is Cl; and R₄ is CF₃.

In one aspect, Z is

Q is

and R₅₁ and R₅₂ are each H. In another aspect, one of R₅₁ and R₅₂ is H,and the other is F or Cl. In a further aspect, one of R₅₁ and R₅₂ is H,and the other is F. In a further aspect, X is CH; Y is CR₄; and R₄ isCF₃.

Representative compounds of the present invention for use in treating orpreventing fibrosis include the compounds listed in Table 1 above andTable 2 below.

TABLE 2 Cmpd # Chemical Structure A1

A2

A3

A4

A5

A6

A7

In one aspect, a compound of the present invention inhibits the activityof one or more αv integrins (e.g., αvβ3, αvβ5, αvβ6, and αvβ8). In afurther aspect, a compound of the present invention inhibits theactivity of αvβ3. In another further aspect, a compound of the presentinvention inhibits the activity of αvβ5. In another further aspect, acompound of the present invention inhibits the activity of αvβ6. Inanother further aspect, a compound of the present invention inhibits theactivity of αvβ8. In yet another further aspect, a compound of thepresent invention inhibits the activity of αvβ3 and αvβ5. In yet anotherfurther aspect, a compound of the present invention inhibits theactivity of αvβ6 and αvβ8. In a further aspect, a compound of thepresent invention inhibits the activity of αvβ3, αvβ5, αvβ6, and/or αvβ8at a submicromolar concentration, e.g., below 1 μM, 0.8 μM, 0.6 μM, 0.5μM, 0.2 μM, or 0.1 μM.

In one aspect, a compound of the present invention inhibits cellularadhesion to vitronectin through the αv integrin (e.g., αvβ3 and αvβ5) ator below an IC₅₀ of 2.0E-07 M using a human dermal microvascularendothelial cell (HMVEC) assay. In a further aspect, a compound of thepresent invention inhibits cellular adhesion to vitronectin through theαv integrin (e.g., αvβ3 and αvβ5) at or below an IC₅₀ of 2.5E-08 M usingan HMVEC assay. In a further aspect, a compound of the present inventioninhibits cellular adhesion to vitronectin through the αv integrin (e.g.,αvβ3 and αvβ5) at or below an IC₅₀ of 1.0E-08 M using an HMVEC assay. Inone aspect, a compound of the present invention inhibits cellularadhesion to vitronectin through the αv integrin (e.g., αvβ3 and αvβ5) ator below an IC₅₀ of 2.5E-07 M using a rat lung microvascular endothelialcell (RLMVEC) assay. In a further aspect, a compound of the presentinvention inhibits cellular adhesion to vitronectin through the αvintegrin (e.g., αvβ3 and αvβ5) at or below an IC₅₀ of 3.5E-08 M using anRLMVEC assay. In one aspect, a compound of the present inventioninhibits cellular adhesion to vitronectin through the αv integrin (e.g.,αvβ3 and αvβ5) at or below an IC₅₀ of 2.0E-08 M using a rabbit aorticendothelial cell (RAEC) assay. In a further aspect, a compound of thepresent invention inhibits cellular adhesion to vitronectin through theαv integrin (e.g., αvβ3 and αvβ5) at or below an IC₅₀ of 1.0E-08 M usingan RAEC assay.

In one aspect, a compound of the present invention inhibits cellularadhesion to fibronectin through the αv integrin (e.g., αvβ6 and αvβ8) ata micromolar concentration (e.g., at or below an IC₅₀ of 1.0E-05 M usinga fibronectin binding assay). In a further aspect, a compound of thepresent invention inhibits cellular adhesion to fibronectin through theαv integrin (e.g., αvβ6 and αvβ8) at a submicromolar concentration(e.g., at or below an IC₅₀ of 1.0E-06 M using a fibronectin bindingassay). In one aspect, a compound of the present invention inhibitscellular adhesion to fibronectin through the αv integrin (e.g., αvβ6 andαvβ8) at a nanomolar concentration (e.g., at or below an IC₅₀ of 1.0E-08M using a fibronectin binding assay). In a further aspect, a compound ofthe present invention inhibits cellular adhesion to fibronectin throughthe αv integrin (e.g., αvβ6 and αvβ8) at a subnanomolar concentration(e.g., at or below an IC₅₀ of 1.0E-09 M using a fibronectin bindingassay).

In one aspect, the compounds of the present invention are selective forone αv integrin (e.g., αvβ3, αvβ5, αvβ6, or αvβ8) over other αvintegrins (e.g., αvβ3, αvβ5, αvβ6, or αvβ8). As used herein, “selective”means that a compound, for example a compound of the invention, inhibitsone αv integrin to a greater extent than other αv integrins.

A “selective αv integrin inhibitor” can be identified, for example, bycomparing the ability of a compound to inhibit one αv integrin activityto its ability to inhibit other αv integrins. For example, a compoundmay be assayed for its ability to inhibit αvβ6 activity, as well asαvβ3, αvβ5, and αvβ8 or other αv integrins.

In certain embodiments, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for one αv integrin over other αv integrins asmeasured by IC₅₀). In various embodiments, the compounds of theinvention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.2-foldto 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-fold to50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold, 1.2-fold to1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold,1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-foldto 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold,2-fold to 25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to500-fold, 2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold,5-fold to 50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to1000-fold, 10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold,10-fold to 500-fold, or 10-fold to 1000-fold selectivity for one αvintegrin over other αv integrins. In various embodiments, the compoundsof the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold,1.5-fold to 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, 10-fold to25-fold, 25-fold to 50-fold, 50-fold to 100-fold, or 100-fold to1000-fold selectivity for one αv integrin over other αv integrins. Invarious embodiments, the compounds of the invention exhibit up 1.2-foldto 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for one αv integrinover other αv integrins.

In one embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ3 over the αvβ5, αvβ6, and/or αvβ8 integrin(e.g., as measured by IC₅₀). In various embodiments, the compounds ofthe invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold,1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-foldto 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold, 1.2-fold to1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold,1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-foldto 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold,2-fold to 25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to500-fold, 2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold,5-fold to 50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to1000-fold, 10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold,10-fold to 500-fold, or 10-fold to 1000-fold selectivity for αvβ3 overthe αvβ5, αvβ6, and/or αvβ8 integrin. In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, 10-foldto 25-fold, 25-fold to 50-fold, 50-fold to 100-fold, or 100-fold to1000-fold selectivity for αvβ3 over the αvβ5, αvβ6, and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, or 10-fold to 25-fold selectivity for αvβ3over the αvβ5, αvβ6, and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ5 over the αvβ3, αvβ6, and/or αvβ8 integrin(e.g., as measured by IC₅₀). In various embodiments, the compounds ofthe invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold,1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-foldto 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold, 1.2-fold to1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold,1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-foldto 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold,2-fold to 25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to500-fold, 2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold,5-fold to 50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to1000-fold, 10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold,10-fold to 500-fold, or 10-fold to 1000-fold selectivity for αvβ5 overthe αvβ3, αvβ6, and/or αvβ8 integrin. In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, 10-foldto 25-fold, 25-fold to 50-fold, 50-fold to 100-fold, or 100-fold to1000-fold selectivity for αvβ5 over the αvβ3, αvβ6, and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, or 10-fold to 25-fold selectivity for αvβ5over the αvβ3, αvβ6, and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ6 over the αvβ3, αvβ5, and/or αvβ8 integrin(e.g., as measured by IC₅₀). In various embodiments, the compounds ofthe invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold,1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-foldto 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold, 1.2-fold to1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold,1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-foldto 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold,2-fold to 25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to500-fold, 2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold,5-fold to 50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to1000-fold, 10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold,10-fold to 500-fold, or 10-fold to 1000-fold selectivity for αvβ6 overthe αvβ3, αvβ5, and/or αvβ8 integrin. In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, 10-foldto 25-fold, 25-fold to 50-fold, 50-fold to 100-fold, or 100-fold to1000-fold selectivity for αvβ6 over the αvβ3, αvβ5, and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, or 10-fold to 25-fold selectivity for αvβ6over the αvβ3, αvβ5, and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ6 over the αvβ8 integrin (e.g., as measuredby IC₅₀). In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.2-fold to 5-fold,1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-fold to 50-fold, 1.2-foldto 100-fold, 1.2-fold to 500-fold, 1.2-fold to 1000-fold, 1.5-fold to2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold, 1.5-fold to 25-fold,1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-fold to 500-fold,1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold, 2-fold to25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to 500-fold,2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold, 5-fold to50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to 1000-fold,10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold, 10-fold to500-fold, or 10-fold to 1000-fold selectivity for αvβ6 over the αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for αvβ6 overthe αvβ8 integrin. In various embodiments, the compounds of theinvention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-foldto 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, or 10-fold to 25-foldselectivity for αvβ6 over the αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ8 over the αvβ3, αvβ5, and/or αvβ6 integrin(e.g., as measured by IC₅₀). In various embodiments, the compounds ofthe invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold,1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-foldto 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold, 1.2-fold to1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold,1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-foldto 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold,2-fold to 25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to500-fold, 2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold,5-fold to 50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to1000-fold, 10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold,10-fold to 500-fold, or 10-fold to 1000-fold selectivity for αvβ8 overthe αvβ3, αvβ5, and/or αvβ6 integrin. In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, 10-foldto 25-fold, 25-fold to 50-fold, 50-fold to 100-fold, or 100-fold to1000-fold selectivity for αvβ8 over the αvβ3, αvβ5, and/or αvβ6integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, or 10-fold to 25-fold selectivity for αvβ8over the αvβ3, αvβ5, and/or αvβ6 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for αvβ8 over the αvβ6 integrin (e.g., as measuredby IC₅₀). In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.2-fold to 5-fold,1.2-fold to 10-fold, 1.2-fold to 25-fold, 1.2-fold to 50-fold, 1.2-foldto 100-fold, 1.2-fold to 500-fold, 1.2-fold to 1000-fold, 1.5-fold to2-fold, 1.5-fold to 5-fold, 1.5-fold to 10-fold, 1.5-fold to 25-fold,1.5-fold to 50-fold, 1.5-fold to 100-fold, 1.5-fold to 500-fold,1.5-fold to 1000-fold, 2-fold to 5-fold, 2-fold to 10-fold, 2-fold to25-fold, 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to 500-fold,2-fold to 1000-fold, 5-fold to 10-fold, 5-fold to 25-fold, 5-fold to50-fold, 5-fold to 100-fold, 5-fold to 500-fold, 5-fold to 1000-fold,10-fold to 25-fold, 10-fold to 50-fold, 10-fold to 100-fold, 10-fold to500-fold, or 10-fold to 1000-fold selectivity for αvβ8 over the αvβ6integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for αvβ8 overthe αvβ6 integrin. In various embodiments, the compounds of theinvention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-foldto 2-fold, 2-fold to 5-fold, 5-fold to 10-fold, or 10-fold to 25-foldselectivity for αvβ8 over the αvβ6 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ6 and αvβ8 over the αvβ3 and/or αvβ5integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ6 and αvβ8 over the αvβ3 and/or αvβ5integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ6 and αvβ8 over the αvβ3 and/or αvβ5 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ6and αvβ8 over the αvβ3 and/or αvβ5 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ3 and αvβ5 over the αvβ6 and/or αvβ8integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ3 and αvβ5 over the αvβ6 and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ3 and αvβ5 over the αvβ6 and/or αvβ8 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ3and αvβ5 over the αvβ6 and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ5 and αvβ6 over the αvβ3 and/or αvβ8integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ5 and αvβ6 over the αvβ3 and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ5 and αvβ6 over the αvβ3 and/or αvβ8 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ5and αvβ6 over the αvβ3 and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ3 and αvβ6 over the αvβ5 and/or αvβ8integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ3 and αvβ6 over the αvβ5 and/or αvβ8integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ3 and αvβ6 over the αvβ5 and/or αvβ8 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ3and αvβ6 over the αvβ5 and/or αvβ8 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ3 and αvβ8 over the αvβ5 and/or αvβ6integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ3 and αvβ8 over the αvβ5 and/or αvβ6integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ3 and αvβ8 over the αvβ5 and/or αvβ6 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ3and αvβ8 over the αvβ5 and/or αvβ6 integrin.

In another embodiment, the compounds of the invention exhibit at least1.2-fold, 1.5-fold, 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or100-fold selectivity for each of αvβ5 and αvβ8 over the αvβ3 and/or αvβ6integrin (e.g., as measured by IC₅₀). In various embodiments, thecompounds of the invention exhibit up 1.2-fold to 1.5-fold, 1.2-fold to2-fold, 1.2-fold to 5-fold, 1.2-fold to 10-fold, 1.2-fold to 25-fold,1.2-fold to 50-fold, 1.2-fold to 100-fold, 1.2-fold to 500-fold,1.2-fold to 1000-fold, 1.5-fold to 2-fold, 1.5-fold to 5-fold, 1.5-foldto 10-fold, 1.5-fold to 25-fold, 1.5-fold to 50-fold, 1.5-fold to100-fold, 1.5-fold to 500-fold, 1.5-fold to 1000-fold, 2-fold to 5-fold,2-fold to 10-fold, 2-fold to 25-fold, 2-fold to 50-fold, 2-fold to100-fold, 2-fold to 500-fold, 2-fold to 1000-fold, 5-fold to 10-fold,5-fold to 25-fold, 5-fold to 50-fold, 5-fold to 100-fold, 5-fold to500-fold, 5-fold to 1000-fold, 10-fold to 25-fold, 10-fold to 50-fold,10-fold to 100-fold, 10-fold to 500-fold, or 10-fold to 1000-foldselectivity for each of αvβ5 and αvβ8 over the αvβ3 and/or αvβ6integrin. In various embodiments, the compounds of the invention exhibitup 1.2-fold to 1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-foldto 5-fold, 5-fold to 10-fold, 10-fold to 25-fold, 25-fold to 50-fold,50-fold to 100-fold, or 100-fold to 1000-fold selectivity for each ofαvβ5 and αvβ8 over the αvβ3 and/or αvβ6 integrin. In variousembodiments, the compounds of the invention exhibit up 1.2-fold to1.5-fold, 1.2-fold to 2-fold, 1.5-fold to 2-fold, 2-fold to 5-fold,5-fold to 10-fold, or 10-fold to 25-fold selectivity for each of αvβ5and αvβ8 over the αvβ3 and/or αvβ6 integrin.

In one aspect, a compound of the present invention inhibits or decreasesformation of blood vessels in a tissue or organ, in vivo or in vitro. Inone aspect, a compound of the present invention decreases the formationof blood vessels below 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or5%, as compared to that in an untreated control. In a further aspect, acompound of the present invention decreases the formation of bloodvessels below 60%, 50%, 40%, 30%, 20%, 10%, or 5%, as compared to thatin an untreated control. In a further aspect, a compound of the presentinvention decreases the formation of blood vessels below 40%, 30%, 20%,10%, or 5%, as compared to that in an untreated control. In one aspect,the tissue is a tissue from the eye, such as a retinal tissue. In oneaspect, the organ is the eye.

In one aspect, a compound of the present invention is efficientlydistributed to the back of the eye, e.g., retina, after topicaladministration. In one aspect, a compound of the present invention isefficiently distributed to the retina within 12 hours, 10 hours, 8hours, 6 hours, 4 hours, 2 hours, or 1 hour, after topicaladministration to the eye. In a further aspect, a compound of thepresent invention is efficiently distributed to the retina within 8hours, 6 hours, 4 hours, 2 hours, or 1 hour, after topicaladministration to the eye.

In one aspect, a compound of the present invention inhibits or decreasesformation of fibrotic tissue in an organ (e.g., kidney, lung, liver, andheart). In one aspect, a compound of the present invention decreases theformation of fibrotic tissue below 90%, 80%, 70%, 60%, 50%, 40%, 30%,20%, 10%, or 5%, as compared to that in an untreated control. In afurther aspect, a compound of the present invention decreases theformation of fibrotic tissue below 60%, 50%, 40%, 30%, 20%, 10%, or 5%,as compared to that in an untreated control. In a further aspect, acompound of the present invention decreases the formation of fibrotictissue below 40%, 30%, 20%, 10%, or 5%, as compared to that in anuntreated control.

Synthesis of the Compounds of the Invention

Compounds of the present invention can be conveniently prepared by avariety of methods familiar to those skilled in the art (e.g., accordingto the methods described in WO 2014/124302, the entire contents of whichare incorporated by reference). The compounds of each of the formulaedescribed herein may be prepared according to the following proceduresfrom commercially available starting materials or starting materialswhich can be prepared using literature procedures. These procedures showthe preparation of representative compounds of this invention. It isunderstood that compounds of the present invention other than thoseillustrated in the following schemes can be made using these schemeswith modifications commonly known in the art (e.g., using differentstarting material, changing reaction solvents, or adjusting reactionduration or temperature).

The compounds of the invention may contain one or more asymmetriccenters and can thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers.Additional asymmetric centers may be present depending upon the natureof the various substituents on the molecule. Each such asymmetric centerwill independently produce two optical isomers. It is intended that allof the possible optical isomers and diastereomers in mixtures and aspure or partially purified compounds are included within the ambit ofthe invention. The invention is meant to comprehend all such isomericforms of these compounds.

The independent syntheses of these diastereomers or theirchromatographic separations may be achieved as known in the art byappropriate modification of the methodology disclosed herein. Theirabsolute stereochemistry may be determined by the x-ray crystallographyof crystalline products or crystalline intermediates which arederivatized, if necessary, with a reagent containing an asymmetriccenter of known absolute configuration.

If desired, racemic mixtures of the compounds may be separated so thatthe individual enantiomers are isolated. The separation can be carriedout by methods well known in the art, such as contacting a racemicmixture of compounds with an enantiomerically pure compound to form adiastereomeric mixture, followed by separation of the individualdiastereomers by standard methods, such as fractional crystallization orchromatography. The diastereomeric mixture is often a mixture ofdiastereomeric salts formed by contacting a racemic mixture of compoundswith an enantiomerically pure acid or base. The diastereomericderivatives may then be converted to the pure enantiomers by cleavage ofthe added chiral residue. The racemic mixture of the compounds can alsobe separated directly by chromatographic methods utilizing chiralstationary phases, which are well known in the art.

Alternatively, any enantiomer of a compound may be obtained bystereoselective synthesis using optically pure starting materials orreagents of known configuration by methods well known in the art.

Some of the compounds of the invention may exist in unsolvated as wellas solvated forms such as, for example, hydrates.

“Solvate” means a solvent addition form that contains either astoichiometric or non-stoichiometric amounts of the solvent molecules.Some compounds have a tendency to trap a fixed molar ratio of thesolvent molecules in the crystalline solid state, thus forming asolvate. If the solvent is water, the solvate formed is a hydrate. Whenthe solvent is alcohol, the solvate formed is an alcoholate. Hydratesare formed by the combination of one or more molecules of water with oneof the substances (e.g., a compound of the invention) in which the waterretains its molecular state as H₂O, such combination being able to formone or more hydrate. In hydrates, the water molecules are attachedthrough secondary valencies by intermolecular forces, in particularhydrogen bridges. Solid hydrates contain water as so-called crystalwater in stoichiometric ratios, where the water molecules do not have tobe equivalent with respect to their binding state. Examples of hydratesinclude sesquihydrates, monohydrates, dehydrates, and trihydrates.Equally suitable are the hydrates of salts of the compounds of theinvention.

For use in medicine, the salts of the compounds of the invention referto non-toxic “pharmaceutically acceptable salts”. Other salts may,however, be useful in the preparation of the compounds of the inventionor pharmaceutically acceptable salts thereof. Salts encompassed withinthe term “pharmaceutically acceptable salts” refer to non-toxic salts ofthe compounds of the invention which can be prepared by reacting thefree base with a suitable organic or inorganic acid. Representativesalts include the following: acetate, benzenesulfonate, benzoate,bicarbonate, bisulfate, bitartrate, borate, bromide, camsylate,carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate,edisylate, estolate, esylate, fumarate, gluceptate, gluconate,glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate,lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate,methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate,N-methylglucamine ammonium salt, oleate, oxalate, pamottle (embonate),palmitate, pantothenate, phosphate/diphosphate, polygalacturonate,salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate,teoclate, tosylate, triethiodide, and valerate. Furthermore, where thecompounds of the invention carry an acidic moiety, suitablepharmaceutically acceptable salts thereof may include alkali metalsalts, e.g., sodium or potassium salts; alkaline earth metal salts,e.g., calcium or magnesium salts; and salts formed with suitable organicligands, e.g., quaternary ammonium salts which may be derived fromammonia or organic amines, such as, for example, diethylamine,triethylamine, ethyldiisopropylamine, procaine, dibenzylamine,N-methylmorpholine, dihydroabietylamine, or methylpiperidine.

The invention includes within its scope prodrugs of the compounds of theinvention. In general, such prodrugs will be functional derivatives ofthe compounds of the invention which are readily convertible in vivointo the required compound. Thus, in the methods of treatment of theinvention, the term “administering” shall encompass the treatment of thevarious disease and conditions described with the compound specificallydisclosed or with a compound which may not be specifically disclosed,but which converts to the specified compound in vivo afteradministration to the patient. Conventional procedures for the selectionand preparation of suitable prodrug derivatives are described, forexample, in “Design of Prodrugs,” ed. H. Bundgaard, Elsevier, 1985.Metabolites of these compounds include active species produced uponintroduction of compounds of the invention into the biological milieu.

The invention also includes one or more metabolites of a compound of theinvention.

The present invention also comprehends deuterium labeled compounds offormula I or Ia or the compounds listed in Table 1 and Table 2, whereina hydrogen atom is replaced by a deuterium atom. The deuterium labeledcompounds comprise a deuterium atom having an abundance of deuteriumthat is substantially greater than the natural abundance of deuterium,e.g., 0.015%.

The term “deuterium enrichment factor” as used herein means the ratiobetween the deuterium abundance and the natural abundance of adeuterium. In one aspect, a compound of the invention has a deuteriumenrichment factor for each deuterium atom of at least 3500 (52.5%deuterium incorporation at each deuterium atom), at least 4000 (60%deuterium incorporation), at least 4500 (67.5% deuterium incorporation),at least 5000 (75% deuterium), at least 5500 (82.5% deuteriumincorporation), at least 6000 (90% deuterium incorporation), at least6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuteriumincorporation), at least 6600 (99% deuterium incorporation), or at least6633.3 (99.5% deuterium incorporation).

Deuterium labeled compounds can be prepared using any of a variety ofart-recognized techniques. For example, deuterium labeled compounds offormula I or II or the compounds listed in Table 1 can generally beprepared by carrying out the procedures disclosed in the Schemes and/orExamples described herein, by substituting a readily available deuteriumlabeled reagent for a non-deuterium labeled reagent.

A compound of the invention or a pharmaceutically acceptable salt orsolvate thereof that contains the aforementioned deuterium atom(s) iswithin the scope of the invention. Further, substitution with deuterium,i.e., ²H, can afford certain therapeutic advantages resulting fromgreater metabolic stability, for example, increased in vivo half-lifeand/or reduced dosage requirements.

In one aspect, the present invention relates to a method of synthesizinga compound of the invention or a pharmaceutically acceptable salt orsolvate thereof.

Biological Assays of the Invention

Cell Adhesion Assays

The ability of compounds of the invention to block cell adhesion tovitronectin and/or fibronectin may be tested with methods or techniquesknown in the art, for example, the procedure described below.

Adhesion plates preparation: Cell culture plates are coated withvitronectin or fibronectin.

Cell culturing and loading: Exemplary cells (e.g., HMVEC cells, RLMVECcells, and RAEC cells) are used for the compound testing. Cells aregrown and then suspended for testing.

Adhesion assay: Test compounds are added to the cell suspension. Afterincubation, the cells that do not adhere to vitronectin- orfibronectin-coated plates are removed by gentle washing. The number ofthe remaining cells is measured. IC₅₀ values are calculated.

αV/β6/αVβ8-LAP-TGF β1 Binding Assay

Integrins αVβ6/αVβ8 coupled beads are treated with an αVβ6/αVβ8 ligand(e.g., LAP TGF-β1 (LAP1)), and the complex is incubated with a primaryantibody (Ab), which can be labeled for detection (e.g., fluorescentlylabeled), and optionally with a secondary antibody, which can be labeledfor detection (e.g., fluorescently labeled). Reaction between integrincoupled beads and the ligand was considered as the full reaction, andreaction without the ligand or a compound of the disclosure wasconsidered as the blank reaction. The complex is analyzed, e.g., byeither plate reader or Flow Cytometer, to determine modulation ofbinding between αVβ6/αVβ8 and the ligand (e.g., LAP-TGF β1) by thecompounds of the present application.

αV/β3/αV/β5-LAP-TGF β1 Binding Assay

Integrins αVβ3/αVβ5 coupled beads are treated with an αVβ3/αVβ5 ligand(e.g., vitronectin), and the complex is treated with a primary antibody(Ab), which can be labeled for detection (e.g., fluorescently labeled),and optionally with a secondary antibody, which can be labeled fordetection (e.g., fluorescently labeled). Reaction between integrincoupled beads and the ligand was considered as the full reaction, andreaction without the ligand or a compound of the disclosure wasconsidered as the blank reaction. The complex is analyzed, e.g., byeither plate reader or Flow Cytometer, to determine modulation ofbinding between αVβ3/αVβ5 and the ligand (e.g., vitronectin) by thecompounds of the present application.

Anti-Angiogenic Activity Assay

The anti-angiogenic ability of compounds of the invention may be testedwith methods or techniques known in the art, for example, the proceduredescribed below.

Chick chorioallantoic membrane (CAM) is grafted with gelatin spongesimpregnated with the test compounds and VEGF. Untreated CAM receivedonly VEGF.

Albumin is removed from hen eggs and incubated. Grafts are placed ondeveloping CAMs and further incubated. CAMs are then fixed, dissectedand imaged for blood vessel growth.

Distribution in plasma, aqueous humor, vitreous humor, and retina of thecompounds of the invention, and the in vivo safety and efficacy of thecompounds of the invention may be tested using animals afteradministration of the compounds to the animals.

Fibrosis can be generally recognized based on the distinct morphology offibrous tissue in a biopsy of the organ in which fibrosis is suspected.Other means for detecting the presence of fibrosis or developingfibrosis include computerized axial tomography (CAT or CT) scan,ultrasound, magnetic resonance imaging (MRI), and monitoring the levelof one or more serum markers known to be indicative of fibrosis (e.g.,various types of collagens). The precise manner of diagnosing fibrosisalso varies depending on the organ where the fibrotic process takesplace. For instance, biopsies are generally effective for diagnosingfibrosis of most organs, whereas endoscopy involving a fiber opticinstrument (e.g., a sigmoidoscope or a colonoscope) can be a lesstraumatic alternative to detect fibrosis of certain organs such as theintestine.

Biopsy for Detecting Fibrosis

Procedures for obtaining biopsy from a given organ or tissue are known,e.g., through exploratory surgery, or a biopsy needle. Upon obtaining abiopsy, the sample is examined and given a score to indicate thepresence and level of fibrosis in the sample. Frequently used scoringsystems include: the METAVIR scoring system, modified HAI (ISHAK)scoring system, and the Knodell scoring system. The criteria used inscoring are well established and known to those of skilled in the art.

Fibrosis Markers

There are numerous known serum markers whose level can be indicative ofthe presence and/or severity of fibrosis, including hyaluronic acid,laminin, undulin (type IV collagen) pro-peptides from types I, II, andIV collagens, lysyl oxidase, prolyl hydroxylase, lysyl hydroxylase,PIIINP, PICP, collagen VI, tenascin, collagen XIV, laminin P1, TIMP-1,MMP-2, α2 macroglobulin, haptoglobin, gamma glutamyl transpeptidase, γglobulin, total bilirubin, and apolipoprotein A1.

In Vivo Bleomycin Induced Pulmonary Fibrosis Model.

Experimental animals are randomly and prospectively assigned to groups.On day 0 and prior to bleomycin induction, animals are administered thefirst dose of vehicle or a compound of the present disclosure. Followingdosing, all animals are anesthetized. A small diameter cannula isinserted into the trachea and saline or bleomycin is slowly infused intothe lungs. Group 1 serves as an untreated control group and receivessaline only (no bleomycin) on day 0. The other groups receive bleomycinon day 0. Treatments with vehicle (e.g., methylcellulose), positivecontrol (e.g., Pirfenidone), or a compound of the present disclosure areadministered once or twice daily via oral gavage (PO). All animals areweighed and evaluated daily for respiratory distress.

Prior to sacrifice, animals are anesthetized and once the animal isdetermined to be non-responsive a shallow incision is made. The tracheais isolated and a transverse cut is made between tracheal ringsapproximately half-way through the trachea. A tracheotomy is performedby the insertion of a cannula through the incision secured with surgicalsuture to the trachea. Following cannulation, the adapter end of thecannula is attached to the mechanical ventilator. The animal isventilated and following an acclimation period, lung volume isstandardized and each animal undergoes a measure of total respiratoryimpedance.

Pharmaceutical Compositions of the Invention

The present invention relates to pharmaceutical compositions comprisinga compound of the invention as an active ingredient. In one aspect, theinvention provides a pharmaceutical composition comprising at least onecompound of formula I, or a pharmaceutically acceptable salt or solvatethereof and one or more pharmaceutically acceptable carriers orexcipients. In one aspect, the invention provides a pharmaceuticalcomposition comprising at least one compound of formula II, IIIa, IIIb,IVa, or IVb, or a pharmaceutically acceptable salt or solvate thereofand one or more pharmaceutically acceptable carriers or excipients. Inone aspect, the invention provides a pharmaceutical compositioncomprising at least one compound selected from Table 1.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

The compounds of the invention can be formulated for oral administrationin forms such as tablets, capsules (each of which includes sustainedrelease or timed release formulations), pills, powders, granules,elixirs, tinctures, suspensions, syrups and emulsions. The compounds ofthe invention can also be formulated for intravenous (bolus orin-fusion), intraperitoneal, topical (e.g., ocular eye-drop),subcutaneous, intramuscular or transdermal (e.g., patch) administration,all using forms well known to those of ordinary skill in thepharmaceutical arts. For example, compounds of the invention for thetreatment of macular degeneration, DR, DME, or macular edema followingRVO, are formulated for topical administration, for example, in the formof eye-drops.

For topical ocular administration, the compositions are provided asophthalmic formulation comprising a compound of the present invention inconcentration between about 0.01 and about 5 weight percent, preferablybetween about 0.1 and about 5.0 weight percent, more preferably betweenabout 0.5 and about 5.0 weight percent, and most preferably betweenabout 0.8 and about 3.0 weight percent.

The ophthalmic formulation of the present invention may be in the formof an aqueous solution comprising an aqueous vehicle.

The aqueous vehicle component of the ophthalmic formulation may comprisewater and at least one ophthalmically acceptable excipient. Preferably,the aqueous vehicle comprises a solution of the one or moreophthalmically acceptable excipients in water.

Suitable ophthalmically acceptable excipients include those selectedfrom the group consisting of a solubility enhancing agent, chelatingagent, preservative, tonicity agent, viscosity/suspending agent, buffer,and pH modifying agent, and a mixture thereof. Preferably, theophthalmically acceptable excipient is selected from the groupconsisting of a solubility enhancing agent, chelating agent,preservative, tonicity agent, viscosity/suspending agent, and pHmodifying agent, and a mixture thereof.

Any suitable ophthalmically acceptable solubility enhancing agent can beused. Examples of a solubility enhancing agent include cyclodextrin,such as those selected from the group consisting ofhydroxypropyl-β-cyclodextrin, methyl-β-cyclodextrin, randomlymethylated-3-cyclodextrin, ethylated-β-cyclodextrin,triacetyl-β-cyclodextrin, peracetylated-β-cyclodextrin,carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin,2-hydroxy-3-(trimethylammonio)propyl-β-cyclodextrin,glucosyl-β-cyclodextrin, sulphated β-cyclodextrin (S-(3-CD),maltosyl-β-cyclodextrin, β-cyclodextrin sulfobutyl ether,branched-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, randomlymethylated-γ-cyclodextrin, and trimethyl-γ-cyclodextrin, and mixturesthereof. Preferably, solubility enhancing agent includes β-cyclodextrinsulfobutyl ether, hyrdoxypropyl-β-cyclodextrin, sulphated β-cyclodextrin(S-β-CD), and maltosyl-β-cyclodextrin, and mixtures thereof.β-cyclodextrin sulfobutyl ether is a particularly preferred solubilityenhancing agent. The solubility enhancing agent(s) may be added in anamount of about 1 to about 20 wt %, preferably about 1 to about 10 wt %,and more preferably about 5 to about 10 wt %.

Any suitable ophthalmically acceptable chelating agent can be used.Examples of a suitable ophthalmically acceptable chelating agent includethose selected from the group consisting of ethylenediaminetetraaceticacid and metal salts thereof, disodium edetate, trisodium edetate, andtetrasodium edetate, and mixtures thereof. Disodium edetate is aparticularly preferred chelating agent. The chelating agent(s) may beadded in an amount of about 0.001 to about 0.05 wt %, preferably about0.001 to about 0.02 wt %, more preferably about 0.002 to about 0.01 wt%, and most preferably about 0.002 to about 0.005 wt %.

Preferably, the aqueous vehicle includes a preservative. Preferredpreservatives include those selected from the group consisting ofquaternary ammonium salts such as benzalkonium halides (preferablybenzalkonium chloride), chlorhexidine gluconate, benzethonium chloride,cetyl pyridinium chloride, benzyl bromide, phenylmercury nitrate,phenylmercury acetate, phenylmercury neodecanoate, merthiolate,methylparaben, propylparaben, sorbic acid, potassium sorbate, sodiumbenzoate, sodium propionate, ethyl p-hydroxybenzoate, propylaminopropylbiguanide, and butyl-p-hydroxybenzoate, sorbic acid, and mixturesthereof. More preferably, the preservative is a quaternary ammonium saltsuch as benzalkonium halides (preferably benzalkoniurn chloride),chlorhexidine gluconate, benzethonium chloride, cetyl pyridiniumchloride, potassium sorbate, sodium benzoate, ethyl p-hydroxybenzoate,butyl p-hydroxybenzoate, or propylaminopropyl biguanide, or mixturesthereof. Propylaminopropyl biguanide is an especially preferredpreservative. The preservative(s) may be used in an amount of about0.00001 to about 0.0001 wt %, preferably about 0.00001 to about 0.00008wt %, and more preferably about 0.00002 to about 0.00005 wt %.

The aqueous vehicle may also include a tonicity agent to adjust thetonicity (osmotic pressure) in order to achieve an ophthalmicallycompatible formulation. The tonicity agent can be selected from thegroup consisting of a glycol (such as propylene glycol, diethyleneglycol, triethylene glycol), glycerol, dextrose, glycerin, mannitol,potassium chloride, and sodium chloride, and a mixture thereof.Preferably, the tonicity agent is selected from the group consisting ofglycerin, mannitol, potassium chloride, and sodium chloride. Morepreferably mannitol and/or sodium chloride (and most preferably amixture thereof) are employed. The tonicity agent(s) may be used in anamount of about 0.05 to about 8 wt %, preferably about 0.1 to about 6 wt%, more preferably about 0.1 to about 4 wt %, and most preferably about0.2 to about 4 wt %.

When a mixture of mannitol and sodium chloride is used as tonicityagents, preferably the weight ratio of mannitol:sodium chloride is about4:1 to about 15:1, more preferably about 6:1 to about 14:1, or 8:1 toabout 14:1 and particularly about 10:1 to about 12:1. If mannitol aloneis used as the tonicity agent, it is preferably used in an concentrationof about 4.5 to about 6.5 wt %, and more preferably in a concentrationof about 5.0 to about 5.5 wt %. If sodium chloride alone is used as thetonicity agent, it is used in a concentration of about 0.05 to about 8wt %, preferably about 0.1 to about 6 wt %, more preferably about 0.1 toabout 4 wt %, and most preferably about 0.2 to about 4 wt %.

The aqueous vehicle preferably also contains a viscosity/suspendingagent. Suitable viscosity/suspending agents include those selected fromthe group consisting of cellulose derivatives, such as methyl cellulose,ethyl cellulose, hydroxyethylcellulose, polyethylene glycols (such aspolyethylene glycol 300, polyethylene glycol 400), carboxymethylcellulose, hydroxypropylmethyl cellulose, and cross-linked acrylic acidpolymers (carbomers), such as polymers of acrylic acid cross-linked withpolyalkenyl ethers or divinyl glycol (Carbopols—such as Carbopol 934,Carbopol 934P, Carbopol 971, Carbopol 974 and Carbopol 974P), and amixture thereof. In preferred embodiments of the present invention, theviscosity/suspending agent is a carbomer, more preferably Carbopol 974P.The viscosity/suspending agent(s) may be present in an amount of about0.05 to about 2 wt %, preferably 0.1 to about 1 wt %, more preferablyabout 0.2 to about 0.8 wt %, and most preferably about 0.3 to about 0.5wt %.

In order to adjust the formulation to an ophthalmically acceptable pH(typically a pH range of about 5.0 to about 9.0, more preferably about5.5 to about 8.5, particularly about 6.0 to about 8.5, about 7.0 toabout 8.5, about 7.2 to about 7.7, about 7.1 to about 7.9, or about 7.5to about 8.0), the formulation may contain a pH modifying agent. The pHmodifying agent is typically a mineral acid or metal hydroxide base,selected from the group of potassium hydroxide, sodium hydroxide, andhydrochloric acid, and mixtures thereof, and preferably sodium hydroxideand/or hydrochloric acid. These acidic and/or basic pH modifying agentsare added to adjust the formulation to the target ophthalmicallyacceptable pH range. Hence it may not be necessary to use both acid andbase—depending on the formulation, the addition of one of the acid orbase may be sufficient to bring the mixture to the desired pH range.

The aqueous vehicle may also contain a buffering agent to stabilize thepH. When used, the buffer is selected from the group consisting of aphosphate buffer (such as sodium dihydrogen phosphate and disodiumhydrogen phosphate), a borate buffer (such as boric acid, or saltsthereof including disodium tetraborate), a citrate buffer (such ascitric acid, or salts thereof including sodium citrate), andε-aminocaproic acid, and mixtures thereof. The buffer agent(s) may bepresent in an amount of about 0.05 to about 5 wt %, preferably 0.1 toabout 5 wt %, more preferably about 0.2 to about 5 wt %, and mostpreferably about 0.5 to about 5 wt %.

The ophthalmic formulation for topical administration to the eye mayfurther comprise a wetting agent. In any embodiment of the presentinvention the wetting agent is preferably a nonionic wetting agent. Morepreferably, the wetting agent is water soluble or swellable. Mostpreferably the wetting agent is water soluble. “Water soluble” is to beunderstood in the manner used in standard texts such as the “Handbook ofPharmaceutical Excipients” (Raymond C Rowe, Paul J Sheskey and Sian COwen, Fifth Edition, Pharmaceutical Press and American PharmacistsAssociation 2006). Suitable classes of wetting agents include thoseselected from the group consisting of polyoxypropylene-polyoxyethyleneblock copolymers (poloxamers), polyethoxylated ethers of castor oils,polyoxyethylenated sorbitan esters (polysorbates), polymers ofoxyethylated octyl phenol (Tyloxapol), polyoxyl 40 stearate, fatty acidglycol esters, fatty acid glyceryl esters, sucrose fatty esters, andpolyoxyethylene fatty esters, and mixtures thereof.

Specific examples of suitable wetting agents include those selected fromthe group consisting of: polyoxyethylene-polyoxypropylene blockcopolymers (poloxamers) such as: polyoxyethylene (160) polyoxypropylene(30) glycol [Pluronic F68], polyoxyethylene (42) polyoxypropylene (67)glycol [Pluronic P123], polyoxyethylene (54) polyoxypropylene (39)glycol [Pluronic P85], polyoxyethylene (196) polyoxypropylene (67)glycol [Poloxamer 407, Pluronic F127], polyoxyethylene (20)polyoxypropylene (20) glycol [Pluronic L44], polyoxyethylenated sorbitanesters (polysorbates) such as poly(oxyethylene)sorbitan monopalmitate(polysorbate 40), poly(oxyethylene)sorbitan monostearate (polysorbate60), poly(oxyethylene)sorbitan tri stearate (polysorbate 65),poly(oxyethylene) sorbitan monooleate (polysorbate 80),poly(oxyethylene) sorbitan monolaurate, poly(oxyethylene) sorbitantrioleate, polyethoxylated ethers of castor oils such as polyoxyethylenehydrogenated castor oil 10, polyoxyethylene hydrogenated castor oil 40,polyoxyethylene hydrogenated castor oil 50 and polyoxyethylenehydrogenated castor oil 60, polyoxyl 40 stearate, sucrose fatty esters,and polyoxyethylene fatty esters, and mixtures thereof.

Preferably, the wetting agent is selected from the group consisting of:polyoxyethylene-polyoxypropylene block copolymers (poloxamers) such as:polyoxyethylene (160) polyoxypropylene (30) glycol [Pluronic F68],polyoxyethylene (42) polyoxypropylene (67) glycol [Pluronic P123],polyoxyethylene (54) polyoxypropylene (39) glycol [Pluronic P85],polyoxyethylene (196) polyoxypropylene (67) glycol [Poloxamer 407,Pluronic F127], and polyoxyethylene (20) polyoxypropylene (20) glycol[Pluronic L44], polyoxyethylenated sorbitan esters (polysorbates) suchas poly(oxyethylene)sorbitan monopalmitate (polysorbate 40),poly(oxyethylene)sorbitan monosteaxate (polysorbate 60),poly(oxyethylene)sorbitan tri stearate (polysorbate 65),poly(oxyethylene) sorbitan monooleate (polysorbate 80),poly(oxyethylene) sorbitan monolaurate, and poly(oxyethylene) sorbitantrioleate and mixtures thereof.

More preferably, the wetting agent is a polyoxyethylene-polyoxypropyleneblock copolymer (poloxamer). Examples of suitable poloxamers include:polyoxyethylene (160) polyoxypropylene (30) glycol [Pluronic F68],polyoxyethylene (42) polyoxypropylene (67) glycol [Pluronic P123],polyoxyethylene (54) polyoxypropylene (39) glycol [Pluronic P85],polyoxyethylene (196) polyoxypropylene (67) glycol [Poloxamer 407,Pluronic F127] and polyoxyethylene (20) polyoxypropylene (20) glycol[Pluronic L44] or a mixture thereof.

Further preferred are wetting agents selected from the group consistingof polyoxyethylene (42) polyoxypropylene (67) glycol [Pluronic PI 23],polyoxyethylene (54) polyoxypropylene (39) glycol [Pluronic P85],polyoxyethylene (196) polyoxypropylene (67) glycol [Poloxamer 407,Pluronic F127] and mixtures thereof.

An especially preferred wetting agent is polyoxyethylene (196)polyoxypropylene (67) glycol [Poloxamer 407, Pluronic F127].

Particularly preferred formulations for topical administration to theeye of the present invention comprise a compound of the presentinvention, a solubility enhancing agent, a cheating agent, apreservative, a tonicity agent, a viscosity/suspending agent, a buffer,and a pH modifying agent. More particularly preferred formulations arecomprised of an aqueous solution of a β-cyclodextrin, a borate salt,boric acid, sodium chloride, disodium edetate, and propylaminopropylbiguanide.

In one aspect, the ophthalmic formulation of the present invention is inthe form of a solution, such as one of the following:

Solution Composition a compound of the invention 0.1-5.0 g a solubilityenhancing agent 1-20 g a buffering agent 0.05-5.0 g an tonicity agent0.05-8 g a chelating agent 1-50 mg a preservative 0.01-0.1 mg water 100ml a compound of the invention 0.8-3.0 g a solubility enhancing agent5-10 g a buffering agent 0.5-5.0 g an tonicity agent 0.2-4 g a chelatingagent 2-5 mg a preservative 0.02-0.05 mg water 100 ml SolutionComposition I II III IV a compound of the invention 2.5 g 2.0 g 1.5 g1.0 g a solubility enhancing agent 10 g 10 g 10 g 5 g buffering agent 11.05 g 1.05 g 1.05 g 1.05 g buffering agent 2 0.285 g 0.285 g 0.285 g0.285 g an tonicity agent 0.25 g 0.25 g 0.25 g 0.25 g a chelating agent2.5 mg 2.5 mg 2.5 mg 2.5 mg a preservative 0.03 mg 0.03 mg 0.03 mg 0.03mg water 100 ml 100 ml 100 ml 100 ml

The ophthalmic formulation of the present invention may also be in theform of a gel or a semi-gel, or both; a jelly; a suspension; anemulsion; an oil; an ointment; a cream; or a spray.

The ophthalmic gel, semi-gel, jelly, suspension, emulsion, oil,ointment, cream, or spray may contain various additives incorporatedordinarily, such as buffering agents (e.g., phosphate buffers, boratebuffers, citrate buffers, tartrate buffers, acetate buffers, aminoacids, sodium acetate, sodium citrate and the like), tonicity agents(e.g., saccharides such as sorbitol, glucose and mannitol, polyhydricalcohols such as glycerin, concentrated glycerin, PEG and propyleneglycol, salts such as sodium chloride), preservatives or antiseptics(e.g., benzalkonium chloride, benzalkonium chloride, P-oxybenzoates suchas methyl p-oxybenzoate or ethyl p-oxybenzoate, benzyl alcohol,phenethyl alcohol, sorbic acid or its salt, thimerosal, chlorobutanoland the like), solubilizing enhancing agents (e.g., cyclodextrins andtheir derivative, water-soluble polymers such as polyvinyl pyrrolidone,surfactants such as tyloxapol, polysorbates), pH modifiers (e.g.,hydrochloric acid, acetic acid, phosphoric acid, sodium hydroxide,potassium hydroxide, ammonium hydroxide and the like), thickening agents(e.g., HEC, hydroxypropyl cellulose, methyl cellulose, HPMC,carboxymethyl cellulose and their salts), chelating agents (e.g., sodiumedetate, sodium citrate, condensed sodium phosphate) and the like. Eachof these additives may be in the amount or concentration similar tothose described for the ophthalmic formulation in the form of a solutionabove.

Furthermore the compounds of the invention may be formulated for topicaladministration by incorporation into novel ophthamlic formulationsincluding but not limited to: microemulsions, liposomes, niosomes, gels,hydrogel, nanoparticles, and nanosuspension.

1. Microemulsions

Microemulsions are dispersion of water and oil facilitated by acombination of surfactant and cosurfactant in a manner to reduceinterfacial tension. These systems are usually characterized by higherthermodynamic stability, small droplet size (approximately 100 nm) andclear appearance. Their transparent appearance is due to the high levelof dispersion of the internal phase, and the size of it ranges from100-1000 angstroms. Processes for forming microemulsions suitable foruse in ophthalmic formulations are described in Vandamne, T. F. ProgRetinal Eye Res 2002; 21:15-34, which is incorporated by reference.

2. Liposomes

Liposomes are lipid vesicles containing aqueous core and have beenwidely exploited in ocular delivery for various drug substances.Depending on the nature of the lipid composition selected, liposomes canprovide extended release of the drug.

3. Niosomes

Niosomes are bilayered structural vesicles made up of nonionicsurfactant and are capable of encapsulating both lipophilic andhydrophilic compounds. They can release the drug independent of pH andenhance ocular bioavailability. Niosomes are microscopic lamellarstructures that are formed on the admixture of nonionic surfactant ofthe alkyl or dialkyl polyglycerol ether class and cholesterol withsubsequent hydration in aqueous media. Structurally niosomes are similarto liposomes, in that they are also made up of a bilayer. However, thebilayer in the case of nisomes is made up of nonionic surface-activeagents rather than phospholipids as in the case of liposomes. Niosomesmay be unilamellar or multilamellar depending on the method used toprepare them. They are capable of entrapping hydrophilic and hydrophobicsolutes. They possess great stability and lack many disadvantagesassociate with liposomes such as high cost and the variable purity ofphospholipids. The properties of niosomes and process for preparing themare well known in the art, see e.g., Wagh, V. D. et al., J Pharm Res2010; 3(7):1558-1563; Kaur, H. et al., Int J Pharm Sci Rev Res 2012;15(1):113-120, each of which is incorporated by reference.

4. Gels

Ophthalmic gels are composed of mucoadhesive polymers that providelocalized delivery of an active ingredient to the eye. Such polymershave a property known as bioadhesion, meaning attachment of a drugcarrier to a specific biological tissue. These polymers are able toextend the contact time of the drug with the biological tissues andthereby improve ocular bioavailability. The choice of the polymer playsa critical role in the release kinetics of the drug from the dosageform. Several bioadhesive polymers are available with varying degree ofmucoadhesive performance. Some examples are carboxymethylcellulose,carbopol, polycarbophil, and sodium alginate. The use of gelformulations in ocular drug deliver has been reviewed in Ali, Y. et al.,Adv Drug Deliv Rev 2006; 58: 1258-1268, which is incorporated byreference.

5. Hydrogels

Hydrogels are three-dimensional, hydrophilic, polymeric networks capableof taking in large amounts of water or biological fluids. Residence timecan be significantly enhanced with a hydrogel formulation. The gelationcan be obtained by changing temperature and pH. Poloxamers, the mostwidely used polymer, contains the hydrophobic part in the centersurrounded by a hydrophilic part. Though they are widely employed toenhance the residence time. Recent perspectives in the use of hydrogelsin ocular drug deliver are described by Gaudana, R., Jwala, J., Boddu,S. H. S., Mitra, A. K. Pharm Res. 2009; 26(5):1197-1216 which isincorporated by reference.

6. Nanoparticles

Nanoparticles are defined as particles with a diameter of less than 1μm, comprising of various biodegradable or non biodegradable polymers,lipids, phospholipids or metals. They can be classified as nanospheresor nanocapsules depending upon whether the drug has been uniformlydispersed or coated within polymeric material. The uptake anddistribution of nanoparticles is dependent on their size. The use ofnanoparticles in ocular drug delivery has recently been reviewed by Hinget al., Int. J. Ophthalmol 2013; 6:390-396, which is incorporated byreference.

7. Nanosuspensions

Nanosuspensions are defined as sub-micron colloidal systems that consistof poorly water soluble drugs suspended in an appropriate dispersionmedium stabilized by surfactants. Usually, nanosuspensions consist ofcolloidal carriers like polymeric resins which are inert in nature.Nanosuspensions enhance drug solubility and thus bioavailability. Unlikemicroemulsions, nanosuspensions are non-irritant. Charge on the surfaceof nanoparticles facilitates their adhesion to the cornea. The use ofnanosuspensions in drug delivery is reviewed in Rabinow, Nature Rev DrugDisc 2004; 785-796, which is incorporated by reference.

The compounds of the present invention can also be administered in theform of a formulation suitable for ocular topical delivery. Detaileddescriptions of formulation suitable for ocular topical delivery aredescribed in Bartlett, J. D. and Jaanus, S. D., Clinical OcularPharmacology, 2008, Elsevier Health Sciences, which is incorporated byreference.

The compounds of the invention may also be coupled with soluble polymersas targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspartamide-phenol, and polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of theinvention may be coupled to a class of biodegradable polymers useful inachieving controlled release of a drug, for example, polylactic acid,polyglycolic acid, copolymers of polylactic and polyglycolic acid,polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters,polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked oramphipathic block copolymers of hydrogels.

The present invention also provides a pharmaceutical compositioncomprising a compound of the invention or a pharmaceutically acceptablesalt or solvate thereof, and a pharmaceutically acceptable carrier orexcipient, and further an active ingredient selected from the groupconsisting of a) an antagonist of integrin α5β1, b) acytotoxic/antiproliferative agent, c) an inhibitor of epidermal-derived,fibroblast-derived, or platelet-derived growth factor, d) an inhibitorof VEGF, e) an inhibitor of Flk-1/KDR, Flt-1, Tck/Tie-2, or Tic-1, andf) an inhibitor of phosphoinositide 3-kinase, and a mixture thereof.

The present invention further provides a pharmaceutical compositioncomprising a compound of the invention or a pharmaceutically acceptablesalt or solvate thereof, and a pharmaceutically acceptable carrier orexcipient, and further an active ingredient selected from the groupconsisting of a) an antagonist of integrin α5β1, b) acytotoxic/antiproliferative agent, c) an inhibitor of epidermal-derived,fibroblast-derived, or platelet-derived growth factors, d) an inhibitorof VEGF, and e) an inhibitor of phosphoinositide 3-kinase, and a mixturethereof.

Nonlimiting examples of antagonists of integrin α5β1 are(S)-2-((R)-2-((S)-2-((S)-2-((S)-1-acetylpyrrolidine-2-carboxamido)-3-(1H-imidazol-5-yl)propanamido)-3-hydroxypropanamido)-3-mercaptopropanamido)succinamide,and JSM6427, described in Stragies, R. et al., J. Med. Chem. 2007,50:3786-3794, herein incorporated by reference.

Nonlimiting examples of cytotoxic/antiproliferative agents are taxol,vincristine, vinblastine, and doxorubicin.

Nonlimiting examples of inhibitors of epidermal-derived,fibroblast-derived, or platelet-derived growth factors are pazopanib,and sunitinib,

Nonlimiting examples of inhibitors of vascular endothelial derivedgrowth factor (VEGF) are bevacizumab and ranibizumab,

Nonlimiting examples of inhibitors of phosphoinositide 3-kinase areindelalisib and 2-morpholin-4-yl-8-phenylchroman-4-one.

Methods of Use

“Fibrosis” refers to a condition involving the development of excessivefibrous connective tissue, e.g., scar tissue, in a tissue or organ. Suchgeneration of scar tissue may occur in response to infection,inflammation, or injury of the organ due to a disease, trauma, chemicaltoxicity, and so on. Fibrosis may develop in a variety of differenttissues and organs, including the liver, kidney, intestine, lung, heart,etc.

Fibrosis of organs or tissues is involved in various diseases ordisorders, such as (1) renal diseases (e.g., tubulointerstitialnephritis), (2) respiratory diseases (e.g., interstitial pneumonia(pulmonary fibrosis)), (3) gastrointestinal diseases (e.g.,hepatocirrhosis, chronic pancreatitis and scirrhous gastric cancer), (4)cardiovascular diseases (myocardial fibrosis), (5) bone and articulardiseases (e.g., bone marrow fibrosis and rheumatoid arthritis), (6) skindiseases (e.g., post surgical scar, burn scar, keloid, hypertrophic scarand scleroderma), (7) obstetric diseases (e.g., hysteromyoma), (8)urologic diseases (prostatic hypertrophy), (9) other diseases (e.g.,Alzheimer's disease, sclerosing peritonitis, type I diabetes and postsurgical adhesion). Accordingly, the tissue fibrosis may be cardiacfibrosis, scleroderma, skeletal muscle fibrosis, hepatic fibrosis,kidney fibrosis, pulmonary fibrosis, intestinal fibrosis, or diabeticfibrosis. For example, a fibrosis may be ongenital hepatic fibrosis(CHF); renal tubulointerstitial fibrosis; pulmonary fibrosis associatedwith an autoimmune disorder (e.g. rheumatoid arthritis, lupus andsarcoidosis); interstitial fibrosis associated with diabeticcardiomyopathy; skeletal muscle fibrosis associated with musculardystrophies (e.g., Becker muscular dystrophy and Duchenne musculardystrophy), denervation atrophies, neuromuscular diseases (e.g., acutepolyneuritis, poliomyelitis, Werdig/Hoffman disease, amyotrophic lateralsclerosis, progressive bulbar atrophy disease), Mediastinal fibrosis(soft tissue of the mediastinum), myelofibrosis (bone marrow),retroperitoneal fibrosis (soft tissue of the retroperitoneum),progressive massive fibrosis (lungs), nephrogenic systemic fibrosis(skin), Crohn's Disease (intestine), Keloid (skin), scleroderma/systemicsclerosis (skin, lungs), arthrofibrosis (knee, shoulder, other joints),Peyronie's disease (penis), dupuytren's contracture (hands or fingers),Some forms of adhesive capsulitis (shoulder).

“Hepatic fibrosis” or “fibrosis of the liver” is the excessiveaccumulation of extracellular matrix proteins including collagen thatoccurs in most types of chronic liver diseases. Advanced liver fibrosisresults in cirrhosis, liver failure, and portal hypertension and oftenrequires liver transplantation. Activated hepatic stellate cells, portalfibroblasts, and myofibroblasts of bone marrow origin have beenidentified as major collagen-producing cells in the injured liver. Thesecells are activated by fibrogenic cytokines such as TGF-β1, angiotensinII, and leptin. The main causes of liver fibrosis in industrializedcountries include chronic alcohol abuse, nonalcoholic steatohepatitis(NASH), iron and copper overload, alcohol-induced liver injury, chronicinfection of hepatitis C, B, and D, hemochromatosis, secondary biliarycirrhosis, NASH, and autoimmune hepatitis.

“Pulmonary fibrosis” or “fibrosis of the lung” is a respiratory diseasein which scars are formed in the lung tissues, leading to seriousbreathing problems. The accumulation of excess fibrous connective tissueleads to thickening of the walls, and causes reduced oxygen supply inthe blood. As a consequence patients suffer from perpetual shortness ofbreath. Pulmonary fibrosis may be a secondary effect of other diseases.Most of these are classified as interstitial lung diseases. Examplesinclude autoimmune disorders, viral infections and bacterial infectionlike tuberculosis which may cause fibrotic changes in both lungs upperor lower lobes and other microscopic injuries to the lung. Idiopathicpulmonary fibrosis can also appear without any known cause. Diseases andconditions that may cause pulmonary fibrosis as a secondary effectinclude: inhalation of environmental and occupational pollutants,hypersensitivity pneumonitis, cigarette smoking, some typical connectivetissue diseases (such as rheumatoid arthritis, SLE and scleroderma),other diseases that involve connective tissue (such as sarcoidosis andWegener's granulomatosis), infections, and certain medications (e.g.,amiodarone, bleomycin (pingyangmycin), busulfan, methotrexate,apomorphine, and nitrofurantoin).

“Cardiac fibrosis” or “fibrosis of the heart” may refer to an abnormalthickening of the heart valves due to inappropriate proliferation ofcardiac fibroblasts, but more commonly refers to the proliferation offibroblasts in the cardiac muscle. Fibrotic cardiac muscle is stifferand less compliant and is seen in the progression to heart failure.Fibrocyte cells normally secrete collagen, and function to providestructural support for the heart. When over-activated this processcauses thickening and fibrosis of the valve, with white tissue buildingup primarily on the tricuspid valve, but also occurring on the pulmonaryvalve. The thickening and loss of flexibility eventually may lead tovalvular dysfunction and right-sided heart failure.

“Renal fibrosis” or “fibrosis of the kidney”, characterized byglomerulosclerosis and tubulointerstitial fibrosis, is the final commonmanifestation of a wide variety of chronic kidney diseases (CKD).Progressive CKD often results in widespread tissue scarring that leadsto the complete destruction of kidney parenchyma and end-stage renalfailure.

Cystic fibrosis (CF) is a genetic disorder that affects mostly the lungsbut also the pancreas, liver, kidneys and intestine. Patients experiencesymptoms including difficulty breathing and coughing up sputum as aresult of frequent lung infections. CF is an autosomal recessivedisorder, caused by mutations in both copies of the gene for the proteincystic fibrosis transmembrane conductance regulator (CFTR). CFTR isinvolved in production of sweat, digestive fluids, and mucus.

A compound of formula Ia modulates (e.g., inhibits the activity of,decreases the expression of, and/or increases the degradation of) afactor (e.g., collagen, TGF-β1) that is involved in the regulation ofthe fibrosis process. For example, a compound of formula Ia is capableof reducing collagen synthesis. In another example, a compound offormula Ia can decrease the production of fibrogenic cytokines (e.g.,TGF-β1). In another example, a compound of formula Ia can reduce theaccumulation of extracellular matrix protein. In yet another example, acompound of formula Ia can inhibit the proliferation of fibroblastcells.

In another example, a compound of formula Ia may inhibit processesmediated by αv integrins. Inhibition and blockade of αvβ6 and/or αvβ8result in a phenotype similar to all of the development effects of lossof TGF-β1 and TGF-β3, suggesting that these integrins are required formost or all important roles of these TGF-β isoforms in development offibrosis. Antagonists of the integrins αvβ6 and/or αvβ8 are thus usefulfor treating and preventing fibrotic activity. For example, TGF-βactivation by the αvβ6 integrin has been shown to play an important rolein models of fibrosis in the lungs, biliary tract, and kidney (Hendersonet al., Nat Med 19, 617 (2013)). The αvβ6 integrin has further beenshown to be overexpressed in human kidney epithelium in membranousglomerulonephritis, diabetes mellitus, IgA nephropathy, Goodpasture'ssyndrome, and Alport syndrome renal epithelium (Am. Journal ofPathology, 2007). In one aspect, a compound of formula Ia treats orprevents fibrosis by inhibiting αvβ6 and/or αvβ8.

Over expression of the αvβ6 integrin has also been shown to play a rolein certain cancers, including but not limited to colorectal carcinomas,thyroid carcinomas, cervical squamous cell carcinomas, and certainbreast carcinomas. Over expression of the αvβ8 integrin has beenassociated with a variety of Th17-drive autoimmune diseases, includingpsoriasis, multiple sclerosis, rheumatoid arthritis, and inflammatorybowel disease. A number of integrin receptors have also been shown toplay a role in foot and mouth disease virus (FMDV).

Thus, in one aspect, the present invention provides a method of treatingor preventing a fibrosis, comprising administering to a subject in needthereof a therapeutically effective amount of a compound of formula Iaor a pharmaceutically acceptable salt or solvate thereof or atherapeutically effective amount of a pharmaceutical composition of theinvention. In one aspect, the invention provides treating a fibrosis. Inone aspect, the invention provides preventing a fibrosis.

In another aspect, the present invention also provides the use of acompound of formula Ia or a pharmaceutically acceptable salt or solvatethereof in the manufacture of a medicament for the treatment orprevention of a fibrosis in a subject. The present invention alsoprovides the use of a compound of formula Ia or a pharmaceuticallyacceptable salt or solvate thereof in treating or preventing a fibrosisin a subject.

In one aspect, the fibrosis is fibrosis of the liver, kidney, intestine,lung, or heart. In a further aspect, the fibrosis is involved in variousdiseases or disorders, such as (1) renal diseases (e.g.,tubulointerstitial nephritis), (2) respiratory diseases (e.g.,interstitial pneumonia (pulmonary fibrosis)), (3) gastrointestinaldiseases (e.g., hepatocirrhosis, chronic pancreatitis and scirrhousgastric cancer), (4) cardiovascular diseases (myocardial fibrosis), (5)bone and articular diseases (e.g., bone marrow fibrosis and rheumatoidarthritis), (6) skin diseases (e.g., post surgical scar, burn scar,keloid, hypertrophic scar and scleroderma), (7) obstetric diseases(e.g., hysteromyoma), (8) urologic diseases (prostatic hypertrophy), (9)other diseases (e.g., Alzheimer's disease, sclerosing peritonitis, typeI diabetes and post surgical adhesion).

Diabetic retinopathy, a closely related condition, is the result ofmicrovascular retinal changes. Hyperglycemia-induced intramural pericytedeath and thickening of the basement membrane lead to incompetence ofthe vascular walls in the retina, which affects the blood-retinalbarrier and makes the retinal blood vessels more permeable. Damagedblood vessels leak fluid and lipids onto the macula, the part of theretina that provides us with detailed vision, causing the macula toswell. Eventually this can progress to develop a condition calledmacular edema.

Accordingly, AMD, DR, DME, and macular edema following central retinalvein occlusion (thrombosis) can be treated or prevented throughadministration (e.g., topical administration) of the compounds orpharmaceutical compositions of the present invention.

The present invention provides a method of treating or preventing adisease or condition in a subject, comprising administering to a subjectin need thereof a therapeutically effective amount of a compound of theinvention or a pharmaceutically acceptable salt or solvate thereof or atherapeutically effective amount of a pharmaceutical composition of theinvention. In one aspect, the invention provides treating a disease orcondition. In one aspect, the invention provides preventing a disease orcondition.

In one aspect, the compound or pharmaceutical composition of theinvention is administered topically. In a further aspect, the compoundor pharmaceutical composition of the invention is administered as anophthalmic solution. In another aspect, the compound or pharmaceuticalcomposition of the invention is administered as an ophthalmic emulsion,suspension, gel, or semi-gel. In another aspect, the compound orpharmaceutical composition of the invention is administered as anophthalmic jelly, oil, ointment, cream, or spray.

The compounds or pharmaceutical compositions of the invention areadministered in dosages effective to inhibit the function of αvβ3, αvβ5,αvβ6 and/or αvβ8 integrins and thus treat or prevent a disease conditionmediated by the αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin.

The present invention provides a method of treating or preventing adisease or condition mediated by an αv integrin in a subject, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula I or a pharmaceutically acceptable saltor solvate thereof or a therapeutically effective amount of apharmaceutical composition a compound of formula I or a pharmaceuticallyacceptable salt or solvate thereof. In one aspect, the disease orcondition is a disease or condition in which angiogenesis is involved.In a further aspect, the disease or condition is a disease or conditionin which ocular angiogenesis is involved.

The present invention also provides a method of treating or preventingan αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin-mediated disease or conditionin a subject, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of formula I or apharmaceutically acceptable salt or solvate thereof or a therapeuticallyeffective amount of a pharmaceutical composition comprising a compoundof formula I or a pharmaceutically acceptable salt or solvate thereof.In one aspect, the disease or condition is a disease or condition inwhich ocular angiogenesis is involved. In one aspect, the disease orcondition is macular degeneration. In one aspect, the disease orcondition is age-related macular degeneration (AMD). In one aspect, thedisease or condition is diabetic retinopathy (DR). In one aspect, thedisease or condition is diabetic macular edema (DME). In one aspect, thedisease or condition is macular edema following retinal vein occlusion(RVO). In one aspect, the condition is fibrosis of the liver, kidney,intestine, lung, and heart. In one aspect, the disease is a renaldisease, a respiratory disease, a gastrointestinal disease, acardiovascular disease, a bone and articular disease, a skin disease, anobstetric disease, or a urologic disease.

The present invention also provides a method of treating or preventingan αvβ3 and/or αvβ5 integrin-mediated disease or condition in a subject,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate thereof or a therapeutically effective amountof a pharmaceutical composition a compound of formula I or apharmaceutically acceptable salt or solvate thereof. In one aspect, thedisease or condition is a disease or condition in which ocularangiogenesis is involved. In one aspect, the disease or condition ismacular degeneration. In one aspect, the disease or condition isage-related macular degeneration (AMD). In one aspect, the disease orcondition is diabetic retinopathy (DR). In one aspect, the disease orcondition is diabetic macular edema (DME). In one aspect, the disease orcondition is macular edema following retinal vein occlusion (RVO).

The present invention provides the use of a compound of formula I or apharmaceutically acceptable salt or solvate thereof in the manufactureof a medicament for the treatment or prevention of a disease orcondition in a subject. The present invention provides the use of acompound of formula I or a pharmaceutically acceptable salt or solvatethereof in treating or preventing a disease or condition in a subject.

The present invention provides the use of a compound of formula I or apharmaceutically acceptable salt or solvate thereof in the manufactureof a medicament for the treatment or prevention of a disease orcondition mediated by an αv integrin in a subject. The present inventionprovides the use of a compound of formula I or a pharmaceuticallyacceptable salt or solvate thereof in treating or preventing a diseaseor condition mediated by an αv integrin in a subject. In one aspect, thedisease or condition is a disease or condition in which angiogenesis isinvolved. In a further aspect, the disease or condition is a disease orcondition in which ocular angiogenesis is involved.

The present invention also provides the use of a compound of formula Ior a pharmaceutically acceptable salt or solvate thereof in themanufacture of a medicament for the treatment or prevention of an αvβ3,αvβ5, αvβ6 and/or αvβ8 integrin-mediated disease or condition in asubject. The present invention provides the use of a compound of formulaI or a pharmaceutically acceptable salt or solvate thereof in treatingor preventing of an αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin-mediateddisease or condition in a subject. In one aspect, the disease orcondition is a disease or condition in which ocular angiogenesis isinvolved. In one aspect, the disease or condition is maculardegeneration. In one aspect, the disease or condition is age-relatedmacular degeneration (AMD). In one aspect, the disease or condition isdiabetic retinopathy (DR). In one aspect, the disease or condition isdiabetic macular edema (DME). In one aspect, the disease or condition ismacular edema following retinal vein occlusion (RVO). In one aspect, thecondition is fibrosis of the liver, kidney, intestine, lung, and heart.In one aspect, the disease is a renal disease, a respiratory disease, agastrointestinal disease, a cardiovascular disease, a bone and articulardisease, a skin disease, an obstetric disease, or a urologic disease.

The present invention provides a compound of formula I or apharmaceutically acceptable salt or solvate thereof for use in treatingor preventing of a disease or condition in a subject.

The present invention provides a compound of formula I or apharmaceutically acceptable salt or solvate thereof for use in treatingor preventing of a disease or condition mediated by an αv integrin in asubject. In one aspect, the disease or condition is a disease orcondition in which angiogenesis is involved. In a further aspect, thedisease or condition is a disease or condition in which ocularangiogenesis is involved.

The present invention also provides a compound of formula I or apharmaceutically acceptable salt or solvate thereof for use in treatingor preventing of an αvβ3, αvβ5, αvβ6 and/or αvβ8 integrin-mediateddisease or condition in a subject. In one aspect, the disease orcondition is a disease or condition in which ocular angiogenesis isinvolved. In one aspect, the disease or condition is maculardegeneration. In one aspect, the disease or condition is age-relatedmacular degeneration (AMD). In one aspect, the disease or condition isdiabetic retinopathy (DR). In one aspect, the disease or condition isdiabetic macular edema (DME). In one aspect, the disease or condition ismacular edema following retinal vein occlusion (RVO). In one aspect, thecondition is fibrosis of the liver, kidney, intestine, lung, and heart.In one aspect, the disease is a renal disease, a respiratory disease, agastrointestinal disease, a cardiovascular disease, a bone and articulardisease, a skin disease, an obstetric disease, or a urologic disease.

Administration of the second therapy in combination typically is carriedout over a defined time period (usually minutes, hours, days or weeksdepending upon the combination selected). “Combination therapy” may be,but generally is not, intended to encompass the administration of two ormore of these therapeutic agents as part of separate monotherapyregimens that incidentally and arbitrarily result in the combinations ofthe present invention. “Combination therapy” is intended to embraceadministration of these therapeutic agents in a sequential manner,wherein each therapeutic agent is administered at a different time, aswell as administration of these therapeutic agents, or at least two ofthe therapeutic agents, in a substantially simultaneous manner.

In accordance with the method of the invention, the individualcomponents of the combination can be administered separately atdifferent times during the course of therapy or concurrently in dividedor single combination forms. The instant invention is therefore to beunderstood as embracing all such regimens of simultaneous or alternatingtreatment, and the term “administering” is to be interpretedaccordingly. It will be understood that the scope of combinations of thecompounds of the invention with other agents useful for treating αvintegrin-mediated conditions includes in principle any combination withany pharmaceutical composition useful for treating fibrosis, maculardegeneration, DR, DME, or macular edema following RVO. When the methodof the invention is a combination treatment of a formulation of thepresent invention topically administered to the eyes and an anti-VEGFprotein or aptamer, the procedures, dosages and frequencies of theanti-VEGF protein or aptamer are as described in the package inserts forthose agents.

The dosage regimen utilizing the compounds of the invention is selectedin accordance with a variety of factors including type, species, age,weight, sex and medical condition of the patient; the severity of thecondition to be treated; and the particular compound or salt thereofemployed. An ordinary skilled physician, veterinarian or clinician canreadily determine and prescribe the effective amount of the drugrequired to prevent, counter or arrest the progress of the condition.

In the methods of the invention, the compounds herein described indetail can form the active ingredient, and are typically administered inadmixture with suitable pharmaceutical diluents, excipients or carriers(collectively referred to herein as “carrier”) suitably selected withrespect to the intended topical administration to the eye and consistentwith conventional pharmaceutical practices.

For purposes of the invention, the following definitions will be used(unless expressly stated otherwise):

“A compound of the invention”, “compounds of the invention”, “a compoundof the present invention”, or “compounds of the present invention”refers to a compound(s) disclosed herein, e.g., a compound(s) of theinvention includes a compound(s) of any of the formulae described hereinincluding formula I and Ia and/or a compound(s) explicitly disclosedherein. Whenever the term is used in the context of the invention it isto be understood that the reference is being made to the free base andthe corresponding pharmaceutically acceptable salts or solvates thereof,provided that such is possible and/or appropriate under thecircumstances.

“Pharmaceutical” or “pharmaceutically acceptable” when used herein as anadjective, means substantially non-toxic and substantiallynon-deleterious to the recipient.

By “pharmaceutical composition” it is further meant that the carrier,diluent, solvent, excipient, and salt must be compatible with the activeingredient of the formulation (e.g., a compound of the invention). It isunderstood by those of ordinary skill in this art that the terms“pharmaceutical formulation” and “pharmaceutical composition” aregenerally interchangeable, and they are so used for the purposes of thisapplication.

“Solution” refers to a clear, homogeneous liquid dosage form thatcontains one or more chemical substances dissolved in a solvent ormixture of mutually miscible solvents. Because molecules of atherapeutic agent substance in solution are uniformly dispersed, the useof solutions as dosage forms generally provides assurance of uniformdosage upon administration and good accuracy when the solution isdiluted or otherwise mixed. “Solution” as disclosed herein contemplatesany variations based on the current state of the art or variationsachieved by one skilled in the art.

“Suspension” refers to a liquid dosage form that contains solidparticles dispersed in a liquid vehicle. “Suspension” as disclosedherein contemplates any variations based on the current state of the artor variations achieved by one skilled in the art.

“Excipient” is used herein to include any other compound that is not atherapeutically or biologically active compound and may be contained inor combined with one or more of the compounds of the present invention.As such, an excipient should be pharmaceutically or biologicallyacceptable or relevant (for example, an excipient should generally benon-toxic to the subject). “Excipient” includes a single such compoundand is also intended to include a plurality of excipients. For thepurposes of the present disclosure the term “excipient” and “carrier”are used interchangeably throughout the description of the presentapplication.

“Therapeutically effective amount” refers to that amount of a drug orpharmaceutical agent that will elicit the biological or medical responseof a tissue, system, animal, or human that is being sought by aresearcher or clinician.

“Treat,” “treating,” or “treatment” refers to decreasing the symptoms,markers, and/or any negative effects of a disease or condition in anyappreciable degree in a subject who currently has the disease orcondition. In some embodiments, treatment may be administered to asubject who exhibits only early signs of a disease or condition for thepurpose of decreasing the risk of developing the disease or condition.In some embodiments, “Treat,” “treating,” or “treatment” refers toamelioration of one or more symptoms of a disease or condition. Forexample, amelioration of one or more symptoms of a disease or conditionincludes a decrease in the severity, frequency, and/or length of one ormore symptoms of a disease or condition.

“Prevent,” “prevention,” or “preventing” refers to any method topartially or completely prevent or delay the onset of one or moresymptoms or features of a disease or condition. Prevention may beadministered to a subject who does not exhibit any sign of a disease orcondition.

“Subject” means a human or animal (in the case of an animal, moretypically a mammal). In one aspect, the subject is a human.

The term “symptom” is defined as an indication of disease, illness,injury, or that something is not right in the body. Symptoms are felt ornoticed by the individual experiencing the symptom, but may not easilybe noticed by others. Others are defined as non-health-careprofessionals.

“αv integrin antagonist” refers to a compound which binds to andinhibits or interferes with the function of one or more of αvβ3, αvβ5,αvβ6, and αvβ8, a compound which binds to and inhibits or interfereswith the function of both αvβ3 and αvβ5 (i.e., a dual αvβ3/αvβ5antagonist), or a compound which binds to and inhibits or interfereswith the function of both αvβ6 and αvβ8 (i.e., a dual αvβ6/αvβ8antagonist). The compounds bind to the receptors as antagonists,blocking or interfering with the binding of the native agonist, such asvitronectin, while not provoking a biological response themselves.

“Bone resorption” refers to the process by which osteoclasts degradebone.

“Alkyl” refers to straight chain or branched alkyl of the number ofcarbon atoms specified (e.g., C₁-C₄ alkyl), or any number within thisrange (methyl, ethyl, propyl, i-propyl, butyl, i-butyl, t-butyl, etc.).

“Alkoxy” refers to straight chain or branched alkoxides of the number ofcarbon atoms specified (e.g., C₁-C₆ alkoxy), or any number within thisrange (methoxy, ethoxy, propoxy, i-propoxy, butoxy, i-butoxy, t-butoxy,etc.).

“Carbocyclic ring” refers to saturated cycloalkyl of the number ofcarbon atoms specified (i.e., C₃ or C₄), such as cyclopropyl andcyclobutyl.

“Heterocyclic ring” refers to saturated heterocyclic ring of the numberof carbon atoms specified (i.e., C₃ or C₄), further comprising oneadditional heteroatoms selected from N, O, and S.

The term “about” refers to a range of values which can be 15%, 10%, 8%,5%, 3%, 2%, 1%, or 0.5% more or less than the specified value. Forexample, “about 10%” can be from 8.5% to 11.5%. In one embodiment, theterm “about” refers to a range of values which are 5% more or less thanthe specified value. In another embodiment, the term “about” refers to arange of values which are 2% more or less than the specified value. Inanother embodiment, the term “about” refers to a range of values whichare 1% more or less than the specified value.

EXAMPLES

Abbreviations used in the following examples and elsewhere herein are:

-   -   AcOH acetic acid    -   Boc₂O di-tert-butyl dicarbonate    -   DCM dichloromethane    -   equiv equivalent(s)    -   EtCO₂H propionic acid    -   EtOAc ethyl acetate    -   EtOH ethanol    -   Et₂O diethyl ether    -   Et₃N triethyl amine    -   hr hour(s)    -   HPLC high-performance liquid chromatography    -   iPrOAc isopropyl acetate    -   iPrMgCl isopropyl magnesium chloride    -   iPr₂NH diisopropyl amine    -   LCMS liquid chromatography-mass spectrometry    -   MeCN acetonitrile    -   n-BuLi n-butyl lithium    -   NMP N-methyl-2-pyrrolidone    -   Pd(OAc)₂ palladium (II) acetate    -   PhMe toluene    -   P(o-tol)₃ tri(O-tolyl)phosphine    -   RT retention time    -   t-BuLi tert-butyl lithium    -   t-BuOK potassium tert-butoxide    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   TLC thin layer chromatography

Example 1. Synthesis of(S)-3-(6-(difluoromethoxy)-pyridine-3-yl)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)propanoic acid (Compound A1)

Compound A1 was made using a convergent synthesis scheme as shown inScheme 2-1: fragment 6b is reacted with fragment 9 to form compound 10,which is further reacted in three steps to form Compound A1.

Synthesis of Fragment 6b (Scheme 1) tert-butyl2-oxopyrrolidine-1-carboxylate (2a)

To a stirred solution of compound 1a (10.0 g, 117 mmol, 1.0 equiv) inCH₂Cl₂, were added (Boc)₂O (25.5 g, 117 mmol, 1.00 equiv) and DMAP(0.022 g, 0.180 mmol, 0.001 equiv) at room temperature and the resultingmixture was stirred for 12 hr. After consumption of the startingmaterial (monitored by TLC), volatiles were removed under reducedpressure to afford compound 2a (19.6 g, 90.3%) as a brown syrup. TLC:50% EtOAc/Hexane (R_(f): 0.40).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 3.74 (t, J=6.8 Hz, 50 (t,J=8.0 Hz, 2H), 2.01 (t, J=7.6 Hz, 2H), 1.52 (s, 9H).

tert-butyl (5-(dimethoxyphosphoryl)-4-oxopentyl)carbamate (3a)

To a stirred solution of iPr₂NH (2.99 mL, 21.8 mmol, 1.35 equiv) in THFand cooled to −10° C., was slowly added hexyl lithium (8.79 mL, 20.0mmol, 1.24 equiv). The reaction mixture was cooled to −60° C.,dimethylmethyl phosphonate (2.20 mL, 20.9 mmol, 1.29 equiv) was addedand the resulting mixture was stirred for 1 h. Then, the temperature wasraised to −40° C., and compound 2a (3.0 g, 16.2 mmol, 1.0 equiv) wasadded and the reaction mixture was stirred for an additional 1 hr. Afterconsumption of the starting material, 2N H₂SO₄ solution (20 mL) wasadded slowly to the reaction and the resulting mixture was stirred at 0°C. for 15 minutes. The aqueous layer was extracted with EtOAc (2×25 mL),and the combined organic extracts were washed with water (25 mL), brine(25 ml), dried over Na₂SO₄, filtered, and evaporated under reducedpressure to afford compound 3a as a brown liquid (5.0 g, crude).

TLC: 80% EtOAc/Hexane (R_(f): 0.30).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 4.85 (brs, 1H, Exc),3.80-3.72 (m, 8H), 3.13-3.07 (m, 2H), 2.67 (t, J=6.8 Hz, 2H), 1.87-1.76(m, 2H), 1.43 (s, 9H).

LC-MS: m/z=308.3 [M+H]⁺ at RT 2.67 (99.1% purity).

tert-butyl (3-(1,8-naphthyridin-2-yl)propyl)carbamate (5a)

To a stirred solution of compound 4a (0.500 g, 4.09 mmol, 1.0 equiv) andcompound 3a (1.26 g, crude, 1.0 equiv) in MeOH (9.17 mL), was added 50%NaOH solution (0.314 mL) and the reaction mixture was stirred at 50° C.for 10 hr. After consumption of the starting material (by TLC), thevolatiles were removed, the crude residue was diluted with EtOAc (15mL), and the organic layer was washed with water (2×15 mL). Theseparated organic layer was dried over Na₂SO₄, filtered, andconcentrated under reduced pressure to afford brown syrup, which waspurified by column chromatography on neutral alumina (80% EtOAc: Hexane)to provide compound 5a (0.980 g, 83.3%) as an off-white solid.

TLC: EtOAc.

NMR Spectroscopy: ¹H NMR (500 MHz, CDCl₃): δ 9.09 (s, 1H), 8.17-8.15 (m,1H), 8.10 (d, J=8.0 Hz, 1H), 7.45 (t, J=8.0 Hz, 1H), 7.41 (t, J=15.0,1H), 4.76 (brs, 1H, Exc), 3.25-3.21 (m, 2H), 3.09 (t, J=10.0 Hz, 2H),2.14-2.08 (m, 2H), 1.42 (s, 9H).

LC-MS: m/z=288 [M−H]⁻ at RT 2.86 (94.7%).

tert-butyl (3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)carbamate(6a)

To a stirred solution of compound 5a (0.25 g, 0.87 mmol, 1.00 equiv) inMeOH (5 mL), Rh/C (catalytic, 5 wt %) was added under N₂ atmosphere andstirred at room temperature for 8 h under hydrogen (balloon pressure)atmosphere. After consumption of the starting material, the reactionmixture was filtered through a pad of CELITE® and the pad was washedwith MeOH (5 mL). The filtrate was evaporated under reduced pressure toafford compound 6a (0.18 g, 71.1%) as a white solid.

TLC: EtOAc.

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 7.05 (d, J=7.6 Hz, 1H),6.34 (d, J=7.2 Hz, 1H), 5.44 (s, 1H), 4.78 (brs, 1H, Exc), 3.41-3.38 (m,2H), 3.16 (d, J=6.0 Hz, 2H), 2.68 (t, J=6.0 Hz, 2H), 2.59 (t, J=7.6 Hz,2H), 1.93-1.81 (m, 4H), 1.44 (s, 9H).

LC-MS: m/z=292.3 [M+H]⁺ at RT 3.41 (97.9% purity).

3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propan-1-amine (6b)

To a stirred solution of 6a (0.25 g, 0.85 mmol, 1.00 equiv) in CH₂Cl₂ (5mL) and cooled to 0° C., was added TFA (0.13 mL, 1.69 mmol, 2.00 equiv).The reaction was warmed to room temperature and then stirred for 4 h.After consumption of the starting material (by TLC), the reactionmixture was concentrated under reduced pressure to afford crude compound6b (0.30 g) as a thick syrup which was used in the next step withoutpurification.

Synthesis of Fragment 9 and Completion of the Synthesis5-bromo-2-(difluoromethoxy)pyridine (2)

To a stirred solution of compound 1 (4.50 g, 25.8 mmol, 1.0 equiv) inanhydrous MeCN (80 mL), was added sodium 2-chloro-2,2-difluoroacetate(4.89 g, 31.0 mmol, 1.20 equiv) at room temperature and resultingmixture was stirred at 70° C. for 48 hr. After consumption of thestarting material (by TLC), the reaction mixture was cooled to roomtemperature and diluted with NH₄Cl solution (30 mL). The aqueous layerwas extracted with EtOAc (2×40 mL). The combined organic layers werewashed with brine solution (2×50 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the crudecompound which was purified by silica gel column chromatography (2%EtOAc/hexane) to afford compound 2 (3.2 g, 57%) as pale yellow syrup.TLC: 5% EtOAc/Hexane (R_(f): 0.5).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 8.25 (d, J=2.8 Hz, 1H),7.82 (dd, J=2.4, 6.4 Hz, 1H), 7.40 (t, J=72.8 Hz, 1H), 6.83 (d, J=8.8Hz, 1H).

LC-MS: m/z=224.7 [M+H]⁺ at RT 4.22 (98.2% purity).

(E)-tert-butyl 3-(6-(difluoromethoxy)pyridin-3-yl)acrylate (3)

To a stirred solution of tert-butyl acrylate (9.99 g, 78.1 mmol, 3.50equiv), Et₃N (8.5 mL, 60.2 mmol, 2.70 equiv), and N-methyl pyrrolidine(20 mL), was added tritolylphosphine (1.17 g, 3.52 mmol, 0.16 equiv)followed by Pd(OAc)₂ (0.50 g, 2.22 mmol, 0.09 equiv). The temperaturewas gradually warmed to 90° C. and compound 2 (5.00 g, 22.3 mmol, 1.0equiv) in NMP (10 mL) was then added drop wise and the resulting mixturewas stirred at 90° C. for 12 hr. After consumption of the startingmaterial (by TLC), the reaction mixture was filtered through a pad ofCELITE® and the pad was washed with EtOAc (50 mL). The filtrate waswashed with cold water (2×50 mL) followed by NaOCl (50 mL), and brinesolution (50 mL). The organic layer was dried over anhydrous Na₂SO₄,filtered, and concentrated under reduced pressure to give the cruderesidue which was purified by silica gel column chromatography (3%EtOAc/hexane) to afford compound 3 (4.0 g, 66%) as yellow solid.

TLC: 5% EtOAc/Hexane (R_(f): 0.5).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 8.28 (d, J=2.4 Hz, 1H),7.88 (dd, J=2.0, 6.4 Hz, 1H), 7.56 (d, J=16.0 Hz, 1H), 7.55 (t, J=45.6Hz, 1H), 6.91 (d, J=8.4 Hz, 1H), 6.34 (d, J=16.0 Hz, 1H), 1.53 (s, 9H).

LC-MS: m/z=272 [M+H]⁺ at RT 4.16 (99.5% purity).

(S)-tert-butyl 3-(benzyl((R)-1-phenylethyl)amino)-3-(6-methoxypyridin-3-yl)propanoate (5)

To a stirred solution of compound 4 (0.39 g, 1.85 mmol, 2.0 equiv) inTHF (5 mL) and cooled to −30° C., was added n-BuLi (0.66 mL, 1.65 mmol,1.79 equiv) and the resulting mixture was then cooled to −78° C.Compound 3 (0.25 g, 0.92 mmol, 1.0 equiv) dissolved in THF (3 mL) wasadded and the reaction mixture was stirred for 30 min and then quenchedwith saturated ammonium chloride. The reaction mixture was extractedwith EtOAc (2×20 mL). The combined organic extracts were washed with 10%AcOH and brine solution, dried over anhydrous Na₂SO₄, filtered, andconcentrated under reduced pressure to give the crude compound (mixtureof 3 and 5, 0.17 g) as thick syrup, which was directly used in the nextstep.

TLC: 5% EtOAc/Hexane (R_(f): 0.5).

LC-MS: m/z=483 [M+H]⁺ at RT 4.66 (75.1% purity).

Synthesis of (s)-tert-butyl3-amino-3-(6-(difluoromethoxy)pyridin-3-yl)propanoate (S-029)

To a stirred solution of compound 5 (0.80 g, crude mixture) in EtOAc (5mL) and AcOH (0.5 mL) under N₂ atmosphere, was added 20% Pd(OH)₂ (50mg). The resulting mixture was then stirred under H₂ atmosphere (40 psi)at room temperature for 2 hr. After consumption of the starting material(monitored by TLC), the reaction mixture was filtered through a pad ofCELITE® and the filtrate was concentrated under reduced pressure toafford crude compound which was purified by silica gel columnchromatography (2% MeOH/CH₂Cl₂) to furnish S-029 (0.3 g, 63%) as yellowsyrup.

TLC: 5% MeOH/CH₂Cl₂ (R_(f): 0.3).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 8.17 (d, J=2.8 Hz, 1H),7.78 (dd, J=2.4, 6.4 Hz, 1H), 7.44 (t, 73.2 Hz, 1H), 6.88 (d, J=8.4 Hz,1H), 4.43-4.40 (m, 1H), 2.65-2.56 (m, 2H), 1.42 (s, 9H).

LC-MS: m/z=274 [M+H]⁺ at RT 2.76 (99.8% purity).

(S,E)-tert-Butyl 3-(6-(tert-butoxy)pyridin-3-yl)-3-((2,2-dimethoxyethylidene)amino)propanoate (7)

To a stirred solution of dimethoxy acetaldehyde (0.44 mL, 2.50 mmol,1.20 equiv, 60% in water) in CH₂Cl₂ (10 mL) and cooled to 0° C., wasadded anhydrous MgSO₄ (10 g) followed by S-029 (600 mg, 2.08 mmol, 1.0equiv) in CH₂Cl₂ (5 mL). The reaction mixture was stirred at roomtemperature for 2 hr and then filtered through a pad of CELITE®. Thefiltrate was concentrated under reduced pressure to afford compound 7(650 mg, crude) as a yellow liquid which was used in the next stepwithout further purification. TLC: 5% MeOH/CH₂Cl₂ (R_(f): 0.5).

(S)-tert-butyl3-(6-(difluoromethoxy)pyridin-3-yl)-3-((2,2-dimethoxyethyl) amino)propanoate (8)

To a stirred solution of compound 7 (0.65 g, crude, 1.0 equiv) in MeOH(7 mL) and cooled to 0° C., was added NaBH(CN)₃ (0.13 g, 2.09 mmol, 1.20equiv) and the resulting mixture was stirred at room temperature for 2hr. After consumption of the starting material (by TLC), the MeOH wasremoved under reduced pressure to give the crude residue which wasdiluted with water (10 mL) and extracted with EtOAc (2×10 ml). Thecombined organic extracts were dried over anhydrous Na₂SO₄, filtered,and concentrated under reduced pressure to give the crude material whichwas purified by silica gel column chromatography (2% MeOH/CH₂Cl₂) toafford compound 8 (0.52 g, 79%) as a thick syrup.

TLC: 5% MeOH/CH₂Cl₂ (R_(f): 0.7).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 8.13 (d, J=2.0 Hz, 1H),7.75 (dd, J=2.4, 6.0 Hz, 1H), 7.44 (t, J=73.2 Hz, 1H), 6.87 (d, J=8.4Hz, 1H), 4.43-4.37 (m, 2H), 4.06-4.02 (m, 1H), 3.60-3.54 (m, 2H), 3.35(s, 3H) 3.31 (s, 3H), 2.66-2.57 (m, 2H), 1.39 (s, 9H).

LC-MS: m/z=377 [M+H]⁺ at RT 2.96 (92.3% purity).

(S)-tert-butyl 3-(6-(difluoromethoxy)pyridin-3-yl)-3-(1-(2,2-dimethoxyethyl)-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)ureido)propanoate(10)

To a stirred solution of compound 8 (375 mg, 0.99 mmol, 1.0 equiv) indry CH₂Cl₂ (5 mL) and cooled to 0° C., was added triphosgene (1.50 mL,2.99 mmol, 3.00 equiv, 20% in PhMe) followed by Et₃N (0.30 mL, 2.09mmol, 2.10 equiv). The reaction mixture was slowly warmed to roomtemperature and then stirred for 2 hr. After consumption of the startingmaterial, the volatiles were evaporated to afford the crude compound 9,which was used directly in the next step without purification.

A solution of compound 9 in DCE (2 mL) was added to a solution ofcompound 6b (400 mg, 1.32 mmol, 1.32 equiv) in CH₂Cl₂ (5 mL) and Et₃N(0.55 mL, 3.98 mmol, 4.00 equiv) at 0° C. and the resulting mixture wasstirred at room temperature for 4 hr. After consumption of the startingmaterial (monitored by TLC), the reaction mixture was concentrated underreduced pressure to give the crude residue which was purified by silicagel column chromatography (2% MeOH/CH₂Cl₂) to afford compound 10 (0.40g, 67%) as a thick syrup.

TLC: 5% MeOH/CH₂Cl₂ (R_(f): 0.2).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 8.13 (d, J=2.8 Hz, 1H),7.79 (dd, J=2.4, 6.4 Hz, 1H), 7.62 (tt, J=72.8 Hz, 1H), 7.12 (d, J=6.4Hz, 1H), 6.86 (d, J=8.4 Hz, 1H), 6.36 (d, J=3.6 Hz, 1H), 6.22 (t, J=4.8Hz, 1H), 5.75 (t, J=7.6 Hz, 1H), 4.26 (t, J=5.2 Hz, 1H), 3.45-3.38 (m,8H), 3.27-3.13 (m, 3H), 2.99-2.93 (m, 2H), 2.71-2.59 (m, 5H), 1.93-1.83(m, 5H), 1.39 (s, 9H).

LC-MS: m/z=594 [M+H]⁺ at RT 3.42 (88.1% purity).

(S)-tert-Butyl 3-(6-(difluoromethoxy)pyridin-3-yl)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)-2,3-dihydro-1H-imidazol-1-yl)propanoate(11)

To a stirred solution of compound 10 (0.20 g, 0.34 mmol, 1.0 equiv) inTHF (4 mL) and at −10° C., was added 1 M sulfuric acid (0.8 mL). Thereaction mixture was slowly warmed to room temperature and then stirredfor 10 hr. After consumption of the starting material (monitored byLCMS), the THF was removed and the crude residue was neutralized withsodium hydroxide (50 wt %) to a pH of −7. The aqueous layer wasextracted with 5% MeOH/CH₂Cl₂ (3×20 mL) and the combined organicextracts were dried over anhydrous Na₂SO₄, filtered, and concentratedunder reduced pressure to furnish compound 11 (0.22 g, crude) as asyrup.

TLC: 10% MeOH/CH₂Cl₂ (R_(f): 0.5).

LC-MS: m/z=530 [M+H]⁺ at RT 4.06 (72.8% purity).

(S)-tert-Butyl 3-(6-(difluoromethoxy)pyridin-3-yl)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)propanoate (12)

To a stirred solution of compound 11 (0.45 g, crude, 1.0 equiv) in EtOH(8 mL) under N₂ atmosphere, was added 20% Pd/C (200 mg). The resultingmixture was stirred under H₂ atmosphere (40 psi) at room temperature for36 hr. After consumption of the starting material the reaction mixturewas filtered through a pad of CELITE®, and the filtrate was concentratedunder reduced pressure to afford crude compound 12, which was purifiedby chiral preparative HPLC to afford compound 12 (450 mg, crude) as anoff-white solid.

TLC: 10% MeOH/CH₂Cl₂ (R_(f): 0.5).

LC-MS: m/z=532.6 [M+H]⁺ at RT 3.99 (80.1% purity).

(S)-3-(6-(difluoromethoxy)pyridin-3-yl)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)propanoic acid (Compound A1)

To a stirred solution of compound 12 (0.40 g, crude, 1.0 equiv) inCH₂Cl₂ (2 mL), cooled to −10° C., and under N₂ atmosphere, was added TFA(0.5 mL). The reaction mixture was slowly warmed to room temperature andthen stirred for 2 hr. After consumption of the starting material, thevolatiles were evaporated to afford crude (400 mg) compound, which waspurified by chiral preparative HPLC to afford compound A1 as anoff-white solid.

TLC: 10% MeOH/CH₂Cl₂ (R_(f): 0.3).

NMR Spectroscopy: ¹H NMR (400 MHz, CD₃OD): δ 8.20 (d, J=2.4 Hz, 1H),7.85 (dd, J=2.4, 6.4 Hz, 1H), 7.53 (t, J=2.4 Hz, 1H), 7.50 (d, J=7.2 Hz,1H), 6.98 (d, J=8.4 Hz, 1H), 6.57 (d, J=7.2 Hz, 1H), 5.51 (dd, J=3.6,8.0 Hz, 1H), 3.68-3.61 (m, 1H), 3.52-3.46 (m, 3H), 3.38 (m, 1H),3.24-3.17 (m, 1H), 3.07-2.98 (m, 2H), 2.90-2.62 (m, 6H), 2.09-1.81 (m,4H).

LC-MS: m/z=476 [M+H]⁺ at RT 2.78 (97.9% purity).

HPLC purity: 96.4%; Chiral Purity: 99%.

The compounds of the present invention described in Examples 2-7 inwhich Z is —CH₂CH₂CH₂— were synthesized using the general reactionscheme shown in Scheme 3. Dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate wasadded to the fluorinated nitrogen heterocycle (Q) aldehyde to form thehept-1-en-3-one. The hept-1-en-3-one was reduced to the correspondinghept-1-en-3-ol using lithium aluminum hydride or sodium borohydride. Thehept-1-en-3ol was then reacted with propionic acid in1,1,1-triethoxyethane and the resulting crude rearrangement product wasreduced with hydrogen and palladium on carbon catalyst to thecorresponding olefin reduction product which was then reacted withaqueous base to form the final nonanoic acid compounds.

Example 2. Synthesis of9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-3-(2-(trifluoromethyl)pyrimidin-5-yl)nonanoicacid (Compound A2)(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(2-(trifluoromethyl)pyrimidin-5-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(3.40 g, 10.0 mmol, 1.00 equiv; Coleman, P. J. et al., J. Med. Chem.2004, 47:4829-4837) in THF (10 mL) at 23° C. and under N₂ atmosphere,was added 2-(trifluoromethyl)pyrimidine-5-carbaldehyde (1.76 g, 10.0mmol, 1.00 equiv) and t-BuOK (1.01 g, 9.00 mmol, 0.900 equiv). Afterstirring for 10 min at 23° C., the reaction mixture was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 2.10g of the title compound (54% yield). NMR Spectroscopy: ¹H NMR (300 MHz,CDCl₃): δ 9.03 (s, 2H), 7.50 (d, J=16.2 Hz, 1H), 7.07 (d, J=7.2 Hz, 1H),6.93 (d, J=16.2 Hz, 1H), 6.35 (d, J=7.2 Hz, 1H), 5.17 (br s, 1H),3.42-3.37 (m, 2H), 2.79-2.64 (m, 4H), 2.62-2.55 (m, 2H), 1.95-1.85 (m,2H), 1.77-1.66 (m, 4H). ¹⁹F NMR (282 MHz, CDCl₃): δ −70.3 (s, 3F).

(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(2-(trifluoromethyl)pyrimidin-5-yl)hept-1-en-3-ol

To(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(2-(trifluoromethyl)pyrimidin-5-yl)hept-1-en-3-one(1.20 g, 3.07 mmol, 1.00 equiv) in THF (15 mL) at −78° C. and under N₂atmosphere, was added LiAlH₄ (1.0 M in THF, 3.07 mL, 3.07 mmol, 1.00equiv). After stirring for 10 min at −78° C., H₂O (116 μL) 15% NaOH aq(116 and H₂O (348 μL) were added sequentially. The reaction mixture wasthen warmed to 23° C. and filtered through a pad of CELITE®. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 560mg of the title compound (46% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.86 (s, 2H), 7.06 (d,J=7.2 Hz, 1H), 6.66 (d, J=16.2 Hz, 1H), 6.53 (dd, J=16.2 Hz, 4.5 Hz,1H), 6.34 (d, J=7.2 Hz, 1H), 4.81 (br s, 1H), 4.50-4.40 (m, 1H),3.42-3.37 (m, 2H), 2.70-2.50 (m, 4H), 1.96-1.40 (m, 8H). ¹⁹F NMR (282MHz, CDCl₃): δ −70.1 (s, 3F).

9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-3-(2-(trifluoromethyl)pyrimidin-5-yl)nonanoicacid (Compound A2)

To(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(2-(trifluoromethyl)pyrimidin-5-yl)hept-1-en-3-ol(560 mg, 1.43 mmol, 1.00 equiv) in MeC(OEt)₃ (14 mL) at 23° C. and underN₂ atmosphere, was added EtCO₂H (107 μL, 1.43 mmol, 1.00 equiv). Afterstirring for 2 hr at 140° C., the reaction mixture was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford acrude rearrangement product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C (103 mg, 0.0969 mmol,6.78 mol %) and H₂ gas was introduced into the reaction mixture with aballoon. After stirring for 1 hr at 23° C., the reaction mixture wasfiltered through a pad of CELITE®. The filtrate was concentrated invacuo to afford a crude olefin reduction product, which was used in thenext step without further purification.

To the above obtained residue in MeOH (10 mL) at 23° C. was added 15%NaOH aq (2.7 mL) under an atmosphere of air. After stirring for 20 minat 60° C., the reaction mixture was neutralized with 3N HCl and thenconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 280mg of the title compound (45% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.79 (s, 2H), 7.24 (d,J=7.2 Hz, 1H), 6.25 (d, J=7.2 Hz, 1H), 3.48-3.40 (m, 2H), 3.38-3.32 (m,1H), 2.75-2.52 (m, 4H), 1.95-1.80 (m, 4H), 1.75-1.58 (m, 4H), 1.40-1.18(m, 6H). ¹⁹F NMR (282 MHz, CDCl₃): δ −70.1 (s, 3F).

Example 3. Synthesis of3-(6-(difluoromethoxy)pyridin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A3) 6-(difluoromethoxy)nicotinaldehyde

To 5-bromo-2-(difluoromethoxy)pyridine (448 mg, 2.00 mmol, 1.00 equiv;Ando, M. et al., Org. Lett. 2006, 8:3805-3808) in THF (10 mL) at −78° C.and under an atmosphere of N₂, was added t-BuLi (1.7 M in pentane, 2.35mL, 4.00 mmol, 2.00 equiv) dropwise over 5 min. After stirring for 20min at −78° C., DMF (0.54 mL, 7.0 mmol, 3.5 equiv) was added to thereaction mixture. After stirring for 20 min at −78° C., 1N HCl aq (10mL) was added and the resulting mixture was warmed to 23° C. The phaseswere separated and the aqueous phase was extracted with EtOAc (3×5 mL).The combined organic phases were washed with brine (10 mL), dried(MgSO₄), and filtered. The filtrate was concentrated in vacuo and theresulting residue was purified by silica gel column chromatographyeluting with hexanes/EtOAc to afford 105 mg of the title compound (30%yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.05 (s, 1H), 8.69 (d,J=2.1 Hz, 1H), 8.24 (dd, J=8.4 Hz, 2.4 Hz, 1H), 7.56 (t, J=72.3 Hz, 1H),7.04 (d, J=8.4 Hz, 1H). ¹⁹F NMR (282 MHz, CDCl₃): δ −89.8 (d, J=72.3 Hz,2F).

(E)-1-(6-(difluoromethoxy)pyridin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(1.57 g, 4.62 mmol, 1.00 equiv) in MeCN (11 mL) at 23° C. and under anatmosphere of N₂, was added 6-(difluoromethoxy)nicotinaldehyde (800 mg,4.62 mmol, 1.00 equiv), LiCl (196 mg, 4.62 mmol, 1.00 equiv) and DBU(0.725 mL, 4.85 mmol, 1.05 equiv). After stirring for 1 hr at 50° C.,the reaction mixture was cooled to 23° C. and then filtered through apad of CELITE®. The filtrate was concentrated in vacuo and the resultingresidue was purified by silica gel column chromatography eluting withCH₂Cl₂/MeOH to afford 1.27 g of the title compound (71% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.32 (d, J=2.4 Hz, 1H),7.92 (dd, J=8.4 Hz, 2.4 Hz, 1H), 7.49 (t, J=72.3 Hz, 1H), 7.47 (d,J=16.2 Hz, 1H), 7.06 (d, J=7.2 Hz, 1H), 6.93 (d, J=8.7 Hz, 1H), 6.70 (d,J=16.2 Hz, 1H), 6.35 (d, J=7.2 Hz, 1H), 4.89 (br s, 1H), 3.42-3.36 (m,2H), 2.76-2.64 (m, 4H), 2.62-2.56 (m, 2H), 1.94-1.85 (m, 2H), 1.80-1.66(m, 4H). ¹⁹F NMR (282 MHz, CDCl₃): δ −89.2 (d, J=72.3 Hz, 2F).

(E)-1-(6-(difluoromethoxy)pyridin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol

To(E)-1-(6-(difluoromethoxy)pyridin-3-yl)-′7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one(1.27 g, 3.28 mmol, 1.00 equiv) in THF (33 mL) at 0° C. and under anatmosphere of N₂, was added LiAlH₄ (1.0 M in THF, 3.28 mL, 3.28 mmol,1.00 equiv). After stirring for 10 min at 0° C., H₂O (124 μL), 15% NaOHaq (124 μL), and H₂O (372 μL) were added sequentially and the resultingmixture was warmed to 23° C. and filtered through a pad of CELITE®. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 1.05g of the title compound (82% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.22 (d, J=2.4 Hz, 1H),7.84 (dd, J=8.4 Hz, 2.4 Hz, 1H), 7.49 (t, J=72.3 Hz, 1H), 7.05 (d, J=7.2Hz, 1H), 6.88 (d, J=8.7 Hz, 1H), 6.66 (d, J=16.2 Hz, 1H), 6.55 (dd,J=16.2 Hz, 4.5 Hz, 1H), 6.33 (d, J=7.2 Hz, 1H), 4.84 (br s, 1H),4.52-4.43 (m, 1H), 3.40-3.37 (m, 2H), 2.72-2.51 (m, 4H), 1.95-1.40 (m,8H). ¹⁹F NMR (282 MHz, CDCl₃): δ −89.0 (d, J=72.5 Hz, 2F).

3-(6-(difluoromethoxy)pyridin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A3)

To(E)-1-(6-(difluoromethoxy)pyridin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol(1.05 g, 2.70 mmol, 1.00 equiv) in MeC(OEt)₃ (27 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (201 μL, 2.70 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was directlyloaded onto silica gel and purified by column chromatography on silicagel eluting with hexanes/EtOAc to afford a crude rearrangement product,which was used in the next step without further purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C (176 mg, 0.165 mmol, 6.11mol %) and H₂ gas was introduced into the reaction mixture with aballoon. After stirring for 1 hr at 23° C., the reaction mixture wasfiltered through a pad of CELITE®. The filtrate was concentrated invacuo to afford a crude olefin reduction product, which was used in thenext step without further purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (4.4 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl andconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 400mg of the title compound (34% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.06 (d, J=2.4 Hz, 1H),7.66 (dd, J=8.4 Hz, 2.4 Hz, 1H), 7.43 (t, J=72.3 Hz, 1H), 7.20 (d, J=8.7Hz, 1H), 6.84 (d, J=7.2 Hz, 1H), 6.25 (d, J=7.2 Hz, 1H), 3.46-3.40 (m,2H), 3.38-3.28 (m, 1H), 2.79-2.40 (m, 4H), 1.95-1.80 (m, 4H), 1.75-1.62(m, 4H), 1.40-1.20 (m, 6H). ¹⁹F NMR (282 MHz, CDCl₃): δ −88.3 (d, J=72.5Hz, 2F).

Example 4. Synthesis of3-(6-fluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A4) 6-fluoroquinoline-3-carbaldehyde

To 2-chloro-6-fluoroquinoline-3-carbaldehyde (2.03 g, 9.68 mmol, 1.00equiv) in DMF (10 mL) at 23° C. and under an atmosphere of N₂, was addedtriethylamine (16.2 mL, 116 mmol, 12.0 equiv), Pd(PPh₃)₄ (559 mg, 0.484mmol, 5.00 mol %), and formic acid (1.29 mL, 34.2 mmol, 5.40 equiv).After stirring for 1 hr at 100° C., the reaction mixture was cooled to23° C. and water (40 mL) and EtOAc (30 mL) were added. The phases wereseparated and the aqueous phase was extracted with EtOAc (3×30 mL). Thecombined organic phases were washed with brine (50 mL), dried (MgSO₄),and filtered. The filtrate was concentrated in vacuo and the residue waspurified by silica gel column chromatography eluting with hexanes/EtOActo afford 734 mg of the title compound (43% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.27 (s, 1H), 9.34 (d,J=2.1 Hz, 1H), 8.60 (d, J=1.8 Hz, 1H), 8.21 (dd, J=9.0 Hz, 4.8 Hz, 1H),7.70-7.60 (m, 2H). ¹⁹F NMR (282 MHz, CDCl₃): δ −110.8 (m, 1F).

(E)-1-(6-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(900 mg, 2.64 mmol, 1.10 equiv) in MeCN (22 mL) at 23° C. and under anatmosphere of N₂, was added 6-fluoroquinoline-3-carbaldehyde (420 mg,2.40 mmol, 1.00 equiv), LiCl (101 mg, 2.40 mmol, 1.00 equiv) and DBU(0.377 mL, 2.52 mmol, 1.05 equiv). After stirring for 1 hr at 75° C.,the reaction mixture was cooled to 23° C. and filtered through a pad ofCELITE®. The filtrate was concentrated in vacuo and the residue waspurified by silica gel column chromatography eluting with CH₂Cl₂/MeOH toafford 900 mg of the title compound (96% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 9.06 (d, J=2.4 Hz, 1H),8.21 (d, J=2.1 Hz, 1H), 8.11 (dd, J=10.6 Hz, 5.7 Hz, 1H), 7.66 (d,J=16.2 Hz, 1H), 7.58-7.43 (m, 2H), 7.06 (d, J=7.2 Hz, 1H), 6.96 (d,J=16.2 Hz, 1H), 6.37 (d, J=7.2 Hz, 1H), 4.76 (br s, 1H), 3.43-3.35 (m,2H), 2.78-2.65 (m, 4H), 2.63-2.56 (m, 2H), 1.94-1.85 (m, 2H), 1.82-1.66(m, 4H). ¹⁹F NMR (282 MHz, CDCl₃): δ −111.9 (m, 1F).

(E)-1-(6-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol

To(E)-1-(6-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one(490 mg, 1.26 mmol, 1.00 equiv) in MeOH (29 mL) at 0° C. and under anatmosphere of air, was added NaBH₄ (71.5 mg, 1.89 mmol, 1.5 equiv).After stirring for 1 hr at 0° C., 1N HCl aq (10 mL) was added and thereaction mixture was then concentrated in vacuo to remove the MeOH. Theresidue was neutralized with NaHCO₃ aq and EtOAc (10 mL) was added. Thephases were separated and the aqueous phase was extracted with EtOAc(3×20 mL). The combined organic phases were washed with brine (30 mL),dried (MgSO₄), and filtered. The filtrate was concentrated in vacuo andthe residue was purified by silica gel column chromatography elutingwith CH₂Cl₂/MeOH to afford 490 mg of the title compound (99% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.95 (s, 1H), 8.06 (dd,J=10.6 Hz, 5.7 Hz, 1H), 7.99 (s, 1H), 7.50-7.40 (m, 2H), 7.06 (d, J=7.2Hz, 1H), 6.75 (d, J=16.2 Hz, 1H), 6.49 (dd, J=16.2 Hz, 4.5 Hz, 1H), 6.34(d, J=7.2 Hz, 1H), 4.94 (br s, 1H), 4.47-4.39 (m, 1H), 3.42-3.38 (m,2H), 2.70-2.47 (m, 4H), 1.96-1.45 (m, 8H). ¹⁹F NMR (282 MHz, CDCl₃): δ−111.8 (m, 1F).

3-(6-fluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A4)

To(E)-1-(6-fluoroquinolin-3-yl)-′7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol(489 mg, 1.25 mmol, 1.00 equiv) in MeC(OEt)₃ (12 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (93.3 μL, 1.25 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford acrude rearrangement product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C (128 mg, 0.121 mmol, 9.68mol %) and H₂ gas was introduced into the reaction with a balloon. Afterstirring for 1 hr at 23° C., the reaction mixture was filtered through apad of CELITE® and the filtrate was concentrated in vacuo to afford acrude olefin reduction product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (3.0 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl andconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 500mg of the title compound (92% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CD₃OD): δ 8.78 (s, 1H), 8.11 (s, 1H),8.00-7.93 (m, 1H), 7.52-7.42 (m, 2H), 7.31 (d, J=7.2 Hz, 1H), 6.35 (d,J=7.2 Hz, 1H), 3.38-3.20 (m, 3H), 2.77-2.42 (m, 4H), 1.90-1.20 (m, 14H).¹⁹F NMR (282 MHz, CD₃OD): δ −110.9 (m, 1F).

Example 5. Synthesis of3-(7-fluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A5)(E)-1-(7-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(749 mg, 2.20 mmol, 1.10 equiv) in MeCN (22 mL) at 23° C. and under anatmosphere of N₂, was added 7-fluoroquinoline-3-carbaldehyde (350 mg,2.00 mmol, 1.00 equiv; Sato, I. et al., Synthesis 2004, 9:1419-1428),LiCl (84.8 mg, 2.00 mmol, 1.00 equiv), and DBU (0.314 mL, 2.10 mmol,1.05 equiv). After stirring for 1 hr at 75° C., the reaction mixture wascooled to 23° C. and then filtered through a pad of CELITE®. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 570mg of the title compound (73% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃\): δ 9.10 (d, J=2.4 Hz, 1H),8.28 (d, J=2.1 Hz, 1H), 7.87 (dd, J=9.0 Hz, 6.0 Hz, 1H), 7.74 (dd, J=9.9Hz, 2.4 Hz, 1H), 7.69 (d, J=16.2 Hz, 1H), 7.42-7.33 (m, 1H), 7.11 (d,J=7.2 Hz, 1H), 6.94 (d, J=16.2 Hz, 1H), 6.37 (d, J=7.2 Hz, 1H), 5.41 (brs, 1H), 3.43-3.37 (m, 2H), 2.78-2.58 (m, 6H), 1.93-1.85 (m, 2H),1.81-1.69 (m, 4H). ¹⁹F NMR (282 MHz, CDCl₃\): δ −107.0 (m, 1F).

(E)-1-(7-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol

To(E)-1-(7-fluoroquinolin-3-yl)-′7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one(300 mg, 0.770 mmol, 1.00 equiv) in MeOH (8 mL) at 0° C. and under anatmosphere of air, was added NaBH₄ (87.4 mg, 2.31 mmol, 3.00 equiv).After stirring for 30 min at 0° C., 1N HCl aq (10 mL) was added and thereaction mixture was concentrated in vacuo to remove the MeOH. Theresulting residue was neutralized with NaHCO₃ aq and EtOAc (10 mL) wasthen added. The phases were separated and the aqueous phase wasextracted with EtOAc (3×20 mL). The combined organic phases were washedwith brine (30 mL), dried (MgSO₄), and filtered. The filtrate wasconcentrated in vacuo and the residue was purified by silica gel columnchromatography eluting with CH₂Cl₂/MeOH to afford 210 mg of the titlecompound (70% yield). NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.98(s, 1H), 8.07 (s, 1H), 7.81 (dd, J=9.0 Hz, 6.0 Hz, 1H), 7.78 (dd, J=9.9Hz, 2.4 Hz, 1H), 7.63 (br s, 1H), 7.39-7.28 (m, 1H), 6.78 (d, J=16.2 Hz,1H), 6.47 (dd, J=16.2 Hz, 4.5 Hz, 1H), 6.36 (d, J=7.2 Hz, 1H), 4.48-4.41(m, 1H), 3.48-3.41 (m, 2H), 2.79-2.67 (m, 4H), 1.97-1.48 (m, 8H). ¹⁹FNMR (282 MHz, CDCl₃): δ −109.9 (m, 1F).

3-(7-fluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A5)

To(E)-1-(7-fluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol(730 mg, 1.71 mmol, 1.00 equiv) in MeC(OEt)₃ (17 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (128 μL, 1.71 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was directlypurified by silica gel column chromatography eluting with hexanes/EtOActo afford a crude rearrangement product, which was used in the next stepwithout further purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C (125 mg, 0.117 mmol, 6.84mol %) and H₂ gas was introduced into the reaction mixture with aballoon. After stirring for 1 hr at 23° C., the reaction mixture wasfiltered through a pad of CELITE®. The filtrate was concentrated invacuo to afford a crude olefin reduction product, which was used in thenext step without further purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (3.0 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl and thenconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 480mg of the title compound (64% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CD₃OD): δ 8.79 (s, 1H), 8.21 (s, 1H),8.00-7.91 (m, 1H), 7.62-7.57 (m, 1H), 7.48-7.38 (m, 2H), 6.47 (d, J=7.2Hz, 1H), 3.48-3.30 (m, 3H), 2.80-2.52 (m, 4H), 1.90-1.20 (m, 14H). ¹⁹FNMR (282 MHz, CD₃OD): δ −111.9 (m, 1F).

Example 6. Synthesis of3-(6,7-difluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A6) 6,7-difluoroquinoline-3-carbaldehyde

To 2-chloro-6,7-difluoroquinoline-3-carbaldehyde (1.44 g, 6.33 mmol,1.00 equiv) in DMF (6.3 mL) at 23° C. and under an atmosphere of N₂, wasadded triethylamine (10.6 mL, 76.0 mmol, 12.0 equiv), Pd(PPh₃)₄ (366 mg,0.317 mmol, 5.00 mol %), and formic acid (1.29 mL, 34.2 mmol, 5.40equiv). After stirring for 1 hr at 100° C., the reaction mixture wascooled to 23° C. and water (30 mL) and EtOAc (20 mL) was added. Thephases were separated and the aqueous phase was extracted with EtOAc(3×20 mL). The combined organic phases were washed with brine (50 mL),dried (MgSO₄), and filtered. The filtrate was concentrated in vacuo andthe residue was purified by silica gel column chromatography elutingwith hexanes/EtOAc to afford 500 mg of the title compound (41% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.26 (s, 1H), 9.35 (d,J=1.2 Hz, 1H), 8.60 (d, J=1.5 Hz, 1H), 7.97 (dd, J=10.8 Hz, 7.5 Hz, 1H),7.97 (dd, J=9.0 Hz, 8.7 Hz, 1H). ¹⁹F NMR (282 MHz, CDCl₃): δ −125.3 (m,1F), −132.3 (m, 1F).

(E)-1-(6,7-difluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(599 mg, 1.76 mmol, 1.10 equiv) in MeCN (5 mL) at 23° C. and under anatmosphere of N₂, was added 6,7-difluoroquinoline-3-carbaldehyde (310mg, 1.60 mmol, 1.00 equiv), LiCl (67.8 mg, 1.60 mmol, 1.00 equiv), andDBU (0.251 mL, 1.68 mmol, 1.05 equiv). After stirring for 1 hr at 75°C., the reaction mixture was cooled to 23° C. and then filtered througha pad of CELITE®. The filtrate was concentrated in vacuo and the residuewas purified by silica gel column chromatography eluting withCH₂Cl₂/MeOH to afford 570 mg of the title compound (84% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 9.07 (d, J=2.4 Hz, 1H),8.20 (d, J=2.1 Hz, 1H), 7.87 (dd, J=10.8 Hz, 7.5 Hz, 1H), 7.66 (d,J=16.2 Hz, 1H), 7.62-7.53 (m, 1H), 7.06 (d, J=7.2 Hz, 1H), 6.93 (d,J=16.2 Hz, 1H), 6.36 (d, J=7.2 Hz, 1H), 4.77 (br s, 1H), 3.43-3.38 (m,2H), 2.79-2.58 (m, 6H), 1.96-1.85 (m, 2H), 1.81-1.69 (m, 4H). ¹⁹F NMR(282 MHz, CDCl₃): δ −129.1 (m, 1F), −133.6 (m, 1F).

(E)-1-(6,7-difluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol

To(E)-1-(6,7-difluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one(1.03 g, 2.53 mmol, 1.00 equiv) in THF (25 mL) at 0° C. and under anatmosphere of N₂, was added LiAlH₄ (1.0 M in THF, 2.53 mL, 2.53 mmol,1.00 equiv). After stirring for 10 min at 0° C., H₂O (96 μL), 15% NaOHaq (96 μL), and H₂O (288 μL) were added sequentially. The reactionmixture was warmed to 23° C. and then filtered through a pad of CELITE®.The filtrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 780mg of the title compound (75% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.95 (d, J=2.4 Hz, 1H),8.00 (d, J=2.1 Hz, 1H), 7.81 (dd, J=10.8 Hz, 7.5 Hz, 1H), 7.52 (d,J=16.2 Hz, 1H), 7.21 (d, J=7.2 Hz, 1H), 6.76 (d, J=16.2 Hz, 1H), 6.48(dd, J=16.2 Hz, 4.5 Hz, 1H), 6.34 (d, J=7.2 Hz, 1H), 4.48-4.42 (m, 1H),3.47-3.41 (m, 2H), 2.79-2.67 (m, 4H), 1.97-1.47 (m, 8H). ¹⁹F NMR (282MHz, CDCl₃): δ −132.1 (m, 1F), −135.1 (m, 1F).

3-(6,7-difluoroquinolin-3-yl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A6)

To(E)-1-(6,7-difluoroquinolin-3-yl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol(780 mg, 1.90 mmol, 1.00 equiv) in MeC(OEt)₃ (19 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (142 μL, 1.90 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford acrude rearrangement product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C (127 mg, 0.119 mmol, 6.26mol %) and H₂ gas was introduced into the reaction with a balloon. Afterstirring for 1 hr at 23° C., the reaction mixture was filtered through apad of CELITE®. The filtrate was concentrated in vacuo to afford a crudeolefin reduction product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (3.2 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl and thenconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 500mg of the title compound (58% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.79 (s, 1H), 7.97 (s, 1H),7.90-7.81 (m, 1H), 7.58-7.47 (m, 1H), 7.24 (d, J=7.2 Hz, 1H), 6.23 (d,J=7.2 Hz, 1H), 3.48-3.32 (m, 3H), 2.80-2.57 (m, 4H), 1.95-1.20 (m, 14H).¹⁹F NMR (282 MHz, CDCl₃): δ −132.3 (m, 1F), −135.5 (m, 1F).

Example 7. Synthesis of9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-3-(7-(trifluoromethyl)quinolin-3-yl)nonanoicacid (Compound A7) 2-chloro-7-iodoquinoline-3-carbaldehyde

To POCl₃ (14.9 mL, 160 mmol, 7.00 equiv) at 0° C. and under anatmosphere of N₂, was added DMF (4.40 mL, 57.1 mmol, 2.50 equiv). Afterstirring for 10 min at 0° C., N-(3-iodophenyl)acetamide (5.96 g, 22.8mmol, 1.00 equiv; Pialat, A. et al., Org. Lett. 2013, 15:1764-1767) wasadded. After stirring for 17 hr at 75° C., the reaction mixture waspoured into ice. The phases were separated and the aqueous phase wasextracted with CH₂Cl₂ (3×50 mL). The combined organic phases were washedwith brine (100 mL), dried (MgSO₄), and filtered. The filtrate wasconcentrated in vacuo and the residue was purified by silica gel columnchromatography eluting with CH₂Cl₂/MeOH to afford 2.9 g of the titlecompound (40% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.55 (s, 1H), 8.72 (s,1H), 8.52 (s, 1H), 7.93 (dd, J=8.4 Hz, 1.5 Hz, 1H), 7.69 (d, J=8.4 Hz,1H).

2-chloro-7-(trifluoromethyl)quinoline-3-carbaldehyde

To 2-chloro-7-iodoquinoline-3-carbaldehyde (2.90 g, 9.13 mmol, 1.00equiv) in DMF (18 mL) at 23° C. and under an atmosphere of N₂, was addedCuI (4.35 g, 22.8 mmol, 2.50 equiv) and FSO₂CF₂CO₂Me (11.6 mL, 91.3mmol, 10.0 equiv). After stirring for 2 hr at 95° C., the reactionmixture was cooled to 23° C. and filtered through a pad of CELITE®. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford1.5 g of the title compound (63% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.60 (s, 1H), 8.82 (s,1H), 8.39 (s, 1H), 8.14 (d, J=8.4 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H). ¹⁹FNMR (282 MHz, CDCl₃): δ −63.2 (s, 3F).

7-(trifluoromethyl)quinoline-3-carbaldehyde

To 2-chloro-7-(trifluoromethyl)quinoline-3-carbaldehyde (1.50 g, 5.78mmol, 1.00 equiv) in DMF (5.8 mL) at 23° C. and under an atmosphere ofN₂, was added triethylamine (9.67 mL, 69.4 mmol, 12.0 equiv), Pd(PPh₃)₄(334 mg, 0.289 mmol, 5.00 mol %), and formic acid (1.18 mL, 31.2 mmol,5.40 equiv). After stirring for 1 hr at 100° C., the reaction mixturewas cooled to 23° C. and water (30 mL) and EtOAc (20 mL) were added. Thephases were separated and the aqueous phase was extracted with EtOAc(3×20 mL). The combined organic phases were washed with brine (50 mL),dried (MgSO₄), and filtered. The filtrate was concentrated in vacuo andthe residue was purified by silica gel column chromatography elutingwith hexanes/EtOAc to afford 412 mg of the title compound (32% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 10.32 (s, 1H), 9.48 (d,J=1.5 Hz, 1H), 8.71 (d, J=1.5 Hz, 1H), 8.51 (s, 1H), 8.15 (d, J=8.4 Hz,1H), 7.86 (d, J=8.4 Hz, 1H). ¹⁹F NMR (282 MHz, CDCl₃): δ −63.1 (s, 3F).

(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(7-(trifluoromethyl)quinolin-3-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(685 mg, 2.01 mmol, 1.10 equiv) in MeCN (9 mL) at 23° C. and under anatmosphere of N₂, was added 7-(trifluoromethyl)quinoline-3-carbaldehyde(412 mg, 1.83 mmol, 1.00 equiv), LiCl (77.6 mg, 1.83 mmol, 1.00 equiv),and DBU (0.287 mL, 1.92 mmol, 1.05 equiv). After stirring for 1 hr at75° C., the reaction mixture was cooled to 23° C. and then filteredthrough a pad of CELITE®. The filtrate was concentrated in vacuo and theresidue was purified by silica gel column chromatography eluting withCH₂Cl₂/MeOH to afford 706 mg of the title compound (88% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 9.19 (d, J=2.4 Hz, 1H),8.42 (d, J=2.1 Hz, 1H), 8.31 (d, J=2.1 Hz, 1H), 8.00 (d, J=9.0 Hz, 1H),7.79 (d, J=9.0 Hz, 1H), 7.69 (d, J=16.2 Hz, 1H), 7.04 (d, J=7.2 Hz, 1H),6.99 (d, J=16.2 Hz, 1H), 6.37 (d, J=7.2 Hz, 1H), 4.78 (br s, 1H),3.41-3.37 (m, 2H), 2.80-2.58 (m, 6H), 1.93-1.85 (m, 2H), 1.81-1.69 (m,4H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.8 (s, 3F).

(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(7-(trifluoromethyl)quinolin-3-yl)hept-1-en-3-ol

To(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(7-(trifluoromethyl)quinolin-3-yl)hept-1-en-3-one(705 mg, 1.60 mmol, 1.00 equiv) in THF (16 mL) at 0° C. and under anatmosphere of N₂, was added LiAlH₄ (1.0 M in THF, 1.60 mL, 1.60 mmol,1.00 equiv). After stirring for 10 min at 0° C., H₂O (54 μL), 15% NaOHaq (54 μL), and H₂O (162 μL) were added sequentially. The reactionmixture was warmed to 23° C. and then filtered through a pad of CELITE®.The filtrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 515mg of the title compound (73% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 9.08 (d, J=2.4 Hz, 1H),8.37 (d, J=2.1 Hz, 1H), 8.08 (d, J=2.1 Hz, 1H), 7.91 (d, J=9.0 Hz, 1H),7.71 (d, J=9.0 Hz, 1H), 7.06 (d, J=7.2 Hz, 1H), 6.79 (d, J=16.2 Hz, 1H),6.53 (dd, J=16.2 Hz, 4.5 Hz, 1H), 6.34 (d, J=7.2 Hz, 1H), 4.89 (br s,1H), 4.48-4.40 (m, 1H), 3.43-3.37 (m, 2H), 2.75-2.57 (m, 4H), 1.97-1.42(m, 8H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.6 (s, 3F).

9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-3-(7-(trifluoromethyl)quinolin-3-yl)nonanoicacid (Compound A7)

To(E)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)-1-(7-(trifluoromethyl)quinolin-3-yl)hept-1-en-3-ol(515 mg, 1.17 mmol, 1.00 equiv) in MeC(OEt)₃ (12 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (87.3 μL, 1.17 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford acrude rearrangement product, which was used in the next step withoutfurther purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added 10% Pd/C 66.6 mg, 0.0626 mmol,5.35 mol %) and H₂ gas was introduced into the reaction mixture with aballoon. After stirring for 1 hr at 23° C., the reaction mixture wasfiltered through a pad of CELITE®. The filtrate was concentrated invacuo to afford a crude olefin reduction product, which was used in thenext step without further purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (4.4 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl andconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with NaHCO₃ aq (2×5 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 300mg of the title compound (53% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.93 (s, 1H), 8.40 (s, 1H),8.03 (s, 1H), 7.91 (d, J=9.0 Hz, 1H), 7.70 (d, J=9.0 Hz, 1H), 7.24 (d,J=7.2 Hz, 1H), 6.23 (d, J=7.2 Hz, 1H), 3.48-3.40 (m, 3H), 2.80-2.59 (m,4H), 1.95-1.20 (m, 14H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.7 (s, 3F).

Example 8. Synthesis of3-(4-chloro-3-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A23)

Compound A23 was made according to the synthesis scheme as shown inScheme 3-1.

(E)-1-(4-chloro-3-(trifluoromethyl)phenyl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one

To dimethyl(2-oxo-6-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hexyl)phosphonate(1.70 g, 5.00 mmol, 1.00 equiv) in THF (10 mL) at 23° C. and under anatmosphere of N₂, was added 4-chloro-3-(trifluoromethyl)benzaldehyde(1.09 g, 5.25 mmol, 1.5 equiv) and t-BuOK (533 mg, 4.75 mmol, 0.950equiv). After stirring for 10 min at 23° C., water (15 mL) and EtOAc (20mL) were added to the reaction mixture. The phases were separated andthe aqueous phase was extracted with EtOAc (3×10 mL). The combinedorganic phases were washed with brine (30 mL), dried (MgSO₄), andfiltered. The filtrate was concentrated in vacuo and the residue waspurified by silica gel column chromatography eluting with CH₂Cl₂/MeOH toafford 1.90 g of the title compound (90% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.81 (s, 1H), 7.61 (d,J=9.1 Hz, 1H), 7.55 (d, J=9.1 Hz, 1H), 7.48 (d, J=16.2 Hz, 1H), 7.04 (d,J=7.2 Hz, 1H), 6.75 (d, J=16.2 Hz, 1H), 6.36 (d, J=7.2 Hz, 1H), 4.96 (brs, 1H), 3.79-3.70 (m, 2H), 3.41-3.37 (m, 2H), 2.73-2.55 (m, 6H),1.97-1.70 (m, 4H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.9 (s, 3F)

(E)-1-(4-chloro-3-(trifluoromethyl)phenyl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol

To(E)-1-(4-chloro-3-(trifluoromethyl)phenyl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-one(1.90 g, 4.50 mmol, 1.00 equiv) in THF (25 mL) at −78° C. and under anatmosphere of N₂, was added LiAlH₄ (1.0 M in THF, 4.5 mL, 4.5 mmol, 1.0equiv). After stirring for 10 min at −78° C., H₂O (171 μL), 15% NaOH aq(171 μL), and H₂O (513 μL) were added sequentially. The reaction mixturewas warmed to 23° C. and then filtered through a pad of celite. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 800mg of the title compound (42% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.66 (s, 1H), 7.46-7.40 (m,2H), 7.07 (d, J=7.2 Hz, 1H), 6.57 (d, J=16.2 Hz, 1H), 6.33 (d, J=7.2 Hz,1H), 6.28 (dd, J=16.2 Hz, 6.0 Hz, 1H), 5.07 (br s, 1H), 4.40-4.30 (m,1H), 3.40-3.33 (m, 2H), 2.82 (br s, 1H), 2.70-2.55 (m, 4H), 1.93-1.40(m, 8H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.6 (s, 3F).

3-(4-chloro-3-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid

To(E)-1-(4-chloro-3-(trifluoromethyl)phenyl)-7-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)hept-1-en-3-ol(800 mg, 1.88 mmol, 1.00 equiv) in MeC(OEt)₃ (19 mL) at 23° C. and underan atmosphere of N₂, was added EtCO₂H (140 μL, 1.88 mmol, 1.00 equiv).After stirring for 2 hr at 140° C., the reaction mixture was directlypurified by silica gel column chromatography eluting with hexanes/EtOActo afford a crude rearrangement product, which was used in the next stepwithout further purification.

To the above obtained residue in MeOH-TFA (10 mL-1 mL) at 23° C. andunder an atmosphere of air, was added Raney®-Nickel (W.R. Grace and Co.Raney® 2800, slurry, in H₂O, active catalyst; 10 drops) and H₂ gas wasintroduced into the reaction with a balloon. After stirring for 3 hr at23° C., the reaction mixture was filtered through a pad of celite. Thefiltrate was concentrated in vacuo to afford a crude olefin reductionproduct, which was used in the next step without further purification.

To the above obtained residue in MeOH (10 mL) at 23° C. and under anatmosphere of air, was added 15% NaOH aq (3.2 mL). After stirring for 20min at 60° C., the reaction mixture was neutralized with 3N HCl and thenconcentrated in vacuo to remove the MeOH. The residual aqueous solutionwas extracted with EtOAc (3×10 mL) and the combined organic phases werewashed with K₂CO₃ aq (2×5 mL), dried (MgSO₄), and filtered. The filtratewas concentrated in vacuo and the residue was purified by silica gelcolumn chromatography eluting with CH₂Cl₂/MeOH to afford 300 mg of thetitle compound (34% yield over 3 steps).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.54 (s, 1H), 7.39 (d,J=9.1 Hz, 1H), 7.33 (d, J=9.1 Hz, 1H), 7.21 (d, J=7.2 Hz, 1H), 6.25 (d,J=7.2 Hz, 1H), 3.51-3.30 (m, 3H), 2.82-2.42 (m, 6H), 1.98-1.20 (m, 12H).¹⁹F NMR (282 MHz, CDCl₃): δ −62.4 (s, 3F).

Example 9. Synthesis of3-(3-chloro-5-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A24)

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.41-7.35 (m, 3H), 7.20 (d,J=7.2 Hz, 1H), 6.23 (d, J=7.2 Hz, 1H), 3.48-3.39 (m, 2H), 3.35-3.22 (m,1H), 2.79-2.48 (m, 6H), 1.95-1.18 (m, 12H). ¹⁹F NMR (282 MHz, CDCl₃): δ−62.7 (s, 3F).

Example 10. Synthesis of3-(3-chloro-5-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A28)

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.51 (d, J=6.3 Hz, 1H),7.47-7.40 (m, 1H), 7.20 (d, J=7.2 Hz, 1H), 7.09 (dd, J=9.0 Hz, 9.0 Hz,1H), 6.24 (d, J=7.2 Hz, 1H), 3.79-3.62 (m, 1H), 3.49-3.39 (m, 2H),2.84-2.49 (m, 6H), 1.98-1.21 (m, 12H). ¹⁹F NMR (282 MHz, CDCl₃): δ −61.8(s, 3F), −111.9 (s, 1H).

Example 11. Synthesis of3-(3-chloro-5-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (Compound A21)

To3-(4-chloro-3-(trifluoromethyl)phenyl)-9-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)nonanoicacid (1) (100 mg, 0.213 mmol, 1.00 equiv) in MeOH (10 mL) at 23° C. andunder an atmosphere of N₂, was added 20% palladium hydroxide on carbon(30 mg, 0.043 mmol, 0.20 equiv) and H₂ gas was introduced into thereaction mixture with a balloon. After stirring for 3 hr at 23° C., thereaction mixture was filtered through a pad of celite. The filtrate wasconcentrated in vacuo and the residue was purified by silica gel columnchromatography eluting with CH₂Cl₂/MeOH to afford 50 mg of the titlecompound (54% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.50-7.30 (m, 4H), 7.25 (d,J=7.2 Hz, 1H), 6.27 (d, J=7.2 Hz, 1H), 3.56-3.41 (m, 2H), 3.37-3.20 (m,1H), 2.79-2.52 (m, 6H), 1.98-1.18 (m, 12H). ¹⁹F NMR (282 MHz, CDCl₃): δ−62.4 (s, 3F).

Example 12. Synthesis of(S)-3-(6-(difluoromethoxy)-pyridine-3-yl)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)propanoic acid (Compound A15s)

Compound A15s was made according to the synthesis scheme as shown inScheme 4.

(E)-tert-butyl 3-(4-chloro-3-(trifluoromethyl)phenyl)acrylate

To 4-bromo-1-chloro-2-(trifluoromethyl)benzene (5.19 g, 20.0 mmol, 1.00equiv) in DMF (10 mL) at 23° C. and under an atmosphere of air, wasadded tert-butyl acrylate (14.7 mL, 100 mmol, 5.00 equiv), Pd(OAc)₂ (269mg, 1.20 mmol, 6.00 mol %), P(o-tol)₃ (730 mg, 2.40 mmol, 12.0 mol %),and Et₃N (8.37 mL, 60.0 mmol, 3.00 equiv). After stirring for 6 hr at110° C., the reaction mixture was cooled to 23° C., filtered, and thefilter cake was rinsed with Et₂O. The combined filtrate wasconcentrated, after which EtOAc (100 mL) and water (100 mL) were addedto the residue. The phases were separated and the organic phase waswashed with water (2×100 mL). The organic phase was dried (MgSO₄) andthe filtrate was concentrated in vacuo. The residue was purified bysilica gel column chromatography eluting with hexanes/EtOAc to afford6.00 g of the title compound (98% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.80 (s, 1H), 7.61-7.43 (m,3H), 6.40 (d, J=16.2 Hz, 1H), 1.53 (s, 9H). ¹⁹F NMR (282 MHz, CDCl₃): δ−63.0 (s, 3F).

(S)-tert-butyl3-(benzyl((R)-1-phenylethyl)amino)-3-(4-chloro-3-(trifluoromethyl)phenyl)propanoate

To (R)—N-benzyl-1-phenylethanamine (6.12 mL, 29.3 mmol, 1.50 equiv) inTHF (90 mL) at 0° C. and under an atmosphere of N₂, was added i-PrMgCl(2.0 M in Et₂O, 29.3 mL, 58.5 mmol, 3.00 equiv). After stirring for 20min at 0° C., the reaction mixture was cooled to −78° C., and(E)-tert-butyl 3-(4-chloro-3-(trifluoromethyl)phenyl)acrylate (5.98 g,19.5 mmol, 1.00 equiv) in THF (20 mL) was added dropwise over 30 min.After stirring for 30 min, 10% AcOH (aq) (100 mL) and Et₂O (200 mL) wereadded and then the reaction mixture was warmed to 23° C. The phases wereseparated and the organic phase was washed with 10% AcOH (aq) (2×200mL). The organic phase was dried (MgSO₄) and the filtrate wasconcentrated in vacuo, after which the residue was purified by silicagel column chromatography eluting with hexanes/EtOAc to afford 7.0 g ofthe title compound (69% yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.71 (s, 1H), 7.61-7.18 (m,12H), 4.44 (t, J=7.2 Hz, 1H), 3.92 (q, J=7.5 Hz, 1H), 3.62 (s, 2H), 2.46(d, J=7.2 Hz, 2H), 1.35-1.26 (m, 3H), 1.26 (s, 9H). ¹⁹F NMR (282 MHz,CDCl₃): δ −62.5 (s, 3F).

(S)-tert-butyl 3-amino-3-(3-(trifluoromethyl)phenyl)propanoate

To (S)-tert-butyl3-(benzyl((R)-1-phenylethyl)amino)-3-(4-chloro-3-(trifluoromethyl)phenyl)propanoate(7.00 g, 13.5 mmol, 1.00 equiv) in MeOH—AcOH (120 mL-12 mL) at 23° C.and under an atmosphere of air, was added 20% Pd(OH)₂/C (1.90 g, 2.70mmol, 20.0 mol %) and H₂ gas was introduced into the reaction mixturewith a balloon. After stirring for 7 hr at 60° C., the reaction mixturewas filtered through a pad of celite. The filtrate was concentrated,after which EtOAc (100 mL) and K₂CO₃ (aq) (100 mL) were added to theresidue. The phases were separated and the aqueous phase was extractedwith EtOAc (2×100 mL). The combined organic phases were washed withbrine (100 mL), dried (MgSO₄), and filtered. The filtrate wasconcentrated in vacuo and the residue was purified by silica gel columnchromatography eluting with hexanes/EtOAc to afford 3.0 g of the titlecompound (77% yield) NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 7.64(s, 1H), 7.60-7.40 (m, 3H), 4.45 (t, J=7.2 Hz, 1H), 2.59 (d, J=7.2 Hz,2H), 1.40 (s, 9H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.5 (s, 3F).

Enhancement of Enantiomeric Purity of (S)-tert-butyl3-amino-3-(3-(trifluoromethyl)phenyl)-propanoate by recrystallization

To (S)-tert-butyl 3-amino-3-(3-(trifluoromethyl)phenyl)propanoate (3.00g, 10.4 mmol, 1.00 equiv) in i-PrOAc (50 mL) at 23° C. and under anatmosphere of air, was added a hot solution of(1R)-(−)-10-camphorsulfonic acid (2.41 g, 10.4 mmol, 1.00 equiv) ini-PrOAc (50 mL). The solution was cooled and kept at 23° C. for 3 hr.The crystals were collected by filtration and dried under vacuum. Theobtained crystals were suspended in CH₂Cl₂ (30 mL) and K₂CO₃ (aq) (30mL) was then added. The phases were separated and the aqueous phase wasextracted with CH₂Cl₂ (2×30 mL). The combined organic phases were washedwith brine (20 mL), dried (MgSO₄), and filtered. The filtrate wasconcentrated in vacuo to afford 2.22 g of the title compound (74%yield).

(S)-tert-butyl3-((2,2-dimethoxyethyl)amino)-3-(3-(trifluoromethyl)phenyl)propanoate

To (S)-tert-butyl 3-amino-3-(3-(trifluoromethyl)phenyl)propanoate (2.22g, 7.67 mmol, 1.00 equiv) at 23° C. and under an atmosphere of air, inTHF (30 mL) was added 2,2-dimethoxyacetaldehyde (60% wt in H₂O, 1.16 mL,7.67 mmol, 1.00 equiv) and sodium triacetoxyborohydride (4.88 g, 23.0mmol, 3.00 equiv). After stirring for 30 min at 23° C., HCl (aq) (50 mL)was added and the reaction mixture was neutralized by the addition ofK₂CO₃. The phases were separated and the aqueous phase was extractedwith EtOAc (3×50 mL). The combined organic phases were washed with brine(100 mL), dried (MgSO₄), and filtered. The filtrate was concentrated invacuo and the residue was purified by silica gel column chromatographyeluting with hexanes/EtOAc to afford 2.00 g of the title compound (69%yield).

NMR Spectroscopy: ¹H NMR (300 MHz, CDCl₃): δ 8.18 (d, J=7.2 Hz, 1H),7.76 (s, 1H), 7.70-7.59 (m, 2H), 4.92-4.85 (m, 1H), 4.71-4.58 (m, 1H),3.59 (dd, J=16.8, 7.2 Hz, 1H), 3.51 (s, 3H), 3.40 (s, 3H), 3.13 (dd,J=16.8, 7.2 Hz, 1H), 2.91 (dd, J=12.3, 4.5 Hz, 1H), 2.75 (dd, J=12.3,4.5 Hz, 1H), 1.36 (s, 9H). ¹⁹F NMR (282 MHz, CDCl₃): δ −62.8 (s, 3F).

(S)-tert-butyl3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)-2,3-dihydro-1H-imidazol-1-yl)-3-(3-(trifluoromethyl)phenyl)propanoate

To triphosgene (629 mg, 2.12 mmol, 0.400 equiv) in THF (10 mL) at 0° C.and under an atmosphere of N₂, was added a solution of (S)-tert-butyl3-((2,2-dimethoxyethyl)amino)-3-(3-(trifluoromethyl)phenyl)propanoate(2.00 g, 5.30 mmol, 1.00 equiv) and triethylamine (2.22 mL, 15.9 mmol,3.00 equiv) in THF (10 mL). After stirring for 30 min at 23° C.,3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propan-1-amine (1.52 g, 7.95mmol, 1.50 equiv) was added. After stirring for 3.0 hr at 40° C., EtOAc(30 mL) and H₂O (20 mL) were added to the reaction mixture. The phaseswere separated and the aqueous phase was extracted with EtOAc (3×20 mL).The combined organic phases were washed with brine (20 mL), dried(MgSO₄), and filtered. The filtrate was concentrated in vacuo to afforda crude urea, which was used in the next step without furtherpurification.

To the above-obtained crude urea in THF (5.5 mL) under an atmosphere ofair, was added 2M H₂SO₄ (aq) (5.5 mL). After stirring for 12 hr at 23°C., K₂CO₃ (aq) (10 mL) was added. The phases were separated and theaqueous phase was extracted with EtOAc (3×15 mL). The combined organicphases were washed with brine (20 mL), dried (MgSO₄), and filtered. Thefiltrate was concentrated in vacuo and the residue was purified bysilica gel column chromatography eluting with CH₂Cl₂/MeOH to afford 1.60g of the title compound (57% yield).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 7.58-7.40 (m, 4H), 7.04 (d,J=7.2 Hz, 1H), 6.32 (d, J=7.2 Hz, 1H), 6.27 (d, J=2.7 Hz, 1H), 6.21 (d,J=2.7 Hz, 1H), 5.72 (dd, J=7.8 Hz, 7.8 Hz, 1H), 4.82 (br s, 1H),3.64-3.58 (m, 2H), 3.40-3.36 (m, 2H), 3.14-2.98 (m, 2H), 2.70-2.50 (m,4H), 2.03-1.80 (m, 4H), 1.38 (s, 9H). ¹⁹F NMR (375 MHz, CDCl₃): δ −62.6(s, 3F).

(S)-tert-butyl3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)-3-(3-(trifluoromethyl)phenyl)propanoate

To (S)-tert-butyl3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)-2,3-dihydro-1H-imidazol-1-yl)-3-(3-(trifluoromethyl)phenyl)propanoate(1.60 g, 3.02 mmol, 1.00 equiv) in MeOH (15 mL) at 23° C. and under anatmosphere of air, was added 20% Pd(OH)₂/C (424 mg, 0.604 mmol, 0.200equiv) and H₂ gas was introduced into the reaction mixture with aballoon. After stirring for 18 hr at 60° C., the reaction mixture wasconcentrated in vacuo to afford 1.6 g of the title compound (99% yield).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 7.59-7.40 (m, 4H), 7.06 (d,J=7.2 Hz, 1H), 6.35 (d, J=7.2 Hz, 1H), 5.52 (dd, J=7.8 Hz, 7.8 Hz, 1H),3.40-3.18 (m, 8H), 3.00-2.82 (m, 2H), 2.70-2.50 (m, 4H), 1.96-1.80 (m,4H), 1.34 (s, 9H). ¹⁹F NMR (375 MHz, CDCl₃): δ −62.5 (s, 3F).

(S)-3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)-3-(3-(trifluoromethyl)phenyl)propanoicacid

To (S)-tert-butyl3-(2-oxo-3-(3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl)imidazolidin-1-yl)-3-(3-(trifluoromethyl)phenyl)propanoate(1.60 g, 3.00 mmol, 1.00 equiv) in CH₂Cl₂ (3 mL) at 23° C. and under anatmosphere of air, was added TFA (3 mL). After stirring for 1 hr at 23°C., the reaction mixture was concentrated in vacuo, after which EtOAc(10 mL) and K₂CO₃ (aq) (10 mL) were added to the residue. The phaseswere separated and the aqueous phase was extracted with EtOAc (3×10 mL).The combined organic phases were washed with brine (10 mL), dried(MgSO₄), and filtered. The filtrate was concentrated in vacuo and theresidue was purified by silica gel column chromatography eluting withCH₂Cl₂/MeOH to afford 400 mg of the title compound (28% yield).

NMR Spectroscopy: ¹H NMR (400 MHz, CDCl₃): δ 7.60 (s, 1H), 7.58-7.40 (m,3H), 7.22 (d, J=7.2 Hz, 1H), 6.27 (d, J=7.2 Hz, 1H), 5.68 (d, J=9.9 Hz,1H), 3.78-3.38 (m, 5H), 3.20-3.08 (m, 1H), 3.00-2.60 (m, 8H), 1.99-1.75(m, 4H). ¹⁹F NMR (375 MHz, CDCl₃): δ −62.5 (s, 3F).

Example 12. Testing of the Compounds of Present Invention in CellAdhesion Assays

The ability of compounds to block adhesion of three primary cellcultures: human dermal microvascular endothelial (HMVEC), rat lungmicrovascular endothelial (RLMVEC), and rabbit aortic endothelial (RAEC)cells, to vitronectin coated plates was determined using the followingprocedure. This test demonstrates inhibition of the interaction of αvintegrin on the cell surface with the ligand, vitronectin.

Adhesion Plates Preparation.

96-well plates were coated with vitronectin in PBS, pH7.4 by incubating50 μL of the solution (10 μg/ml) for 1.5 h at room temperature orovernight at 4° C. The plates then were blocked with 1% BSA in PBS (30min at room temperature) and washed with PBS.

Cell Culturing and Loading.

HMVEC cells (passages p 9-14) (from Lonza, Allendale, N.J.) RLMVEC cells(p 4-14) (from Vec Technology, Rensselaer, N.Y.) and RAEC cells (p 4-14)(from CellBiologics, Chicago, Ill.) were used for the compound testing.Cells were grown in T175 tissue culture flasks and dislodged by gentle 3min treatment with Accutase (Life Technologies). After washing, thecells in suspension in RPMI-1640 (Life Technologies) were loaded withcalcein-AM (5 μM) (Life Technologies) for 30 min at 37° C. andre-suspended into RPMI w/o phenol red medium containing 10% FBS.

Adhesion Assay.

The cell suspension was aliquoted into the wells at a density of 1.0×10⁵cells/well (RLMVEC) and 5.0×10⁴ (HMVEC, and RAEC). The test compoundswere added at the same time with the cells. The plates were incubatedfor 1.5 h at 37° C. The cells that did not adhere during this incubationwere removed by gentle washing. The wash was performed by 2 cycles ofaspiration of the supernatant and addition of 100 μL of the pre-warmedfresh DPBS (Life Technologies). A fluorescence of the remaining cells ismeasured using multimode plate reader (Victor 2V, PerkinElmer) at anexcitation/emission wavelengths of 485/535 nm. The compounds were testedstarting with maximal concentration of 1 μM with half-log dilutionschedule. IC₅₀ values were calculated with Prism 5 (GraphPad, CA) byfixing the bottom of the curves to a value of blank for empty wellsfluorescence.

Example 13. Testing of the Compounds of Present Invention in α_(V)Integrin Binding Assays

All α_(V) Integrins are known to bind to proteins with a RGD motif. TwoRGD ligands were used in this study: Vitronectin (VN) as a ligand forα_(V)β₃ and α_(V)β₅ (Wayner et al., J. Cell Biol., 113 (4), 919-929,1991), and LAP TGF-β1 (LAP1) as a ligand for α_(V)β₆ and α_(V)β₈(Rognoni et al., Nat. Med., 20(4): 350-359, 2014). CWHM12 was used as apositive control for α_(V)β₆ and α_(V)β₈ (Henderson et al., Nat. Med.19(12), 10.1038/nm.3282 2013), and Cilengitide as a positive control forα_(V)β₃ and α_(V)β₅ (Kumar et al., J. Pharmacol. Exp. Ther., 283,843-853, 1997).

The integrin coupled Dyna beads were allowed to interact with respectiveligands. The integrin-ligand complex was detected with eitherPrimary/Secondary Antibody conjugated with Fluorescein Isothiocyanate(FITC). For α_(V)β3 and α_(V)β5, Vitronectin was used as a ligand and aprimary antibody conjugated with FITC (Anti-VN-FITC Ab) was used todetect the interaction. For α_(V)β6 and α_(V)β8, LAP-TGF (31 was used asligand and a primary antibody against LAP1 (Anti-LAP1 Ab) and asecondary antibody conjugated with FITC were used to detect theα_(V)β6/α_(V)β8-LAP-TGF (31 complex. Fluorescence was measured by FlowCytometry analysis.

Activation of Beads.

5 mg of Dyna beads were weighed in a low protein binding microfuge(Eppendorf) tube (1.5 mL volume). The beads were re-suspended in 1 mL ofSodium Phosphate Buffer and vortexed at high speed for 30 seconds. Thetube was then placed in a tube roller and tilt rotated for 10 min. Aftertilt rotation, the tube was placed on the Magna Spin and the beads wereallowed to settle. The supernatant was discarded and the beads werewashed three times. The beads were then re-suspended in 100 μL of SodiumPhosphate Buffer and 20 μL of washed beads were distributed into 5 lowprotein binding Eppendorf tubes (1 mg of beads in each tube). The beadswere used for coupling integrins.

Coupling of Dyna Beads with Integrins.

20 μL of (1 mg) of beads were mixed with 20 μL of integrins (20 μg) and20 μL of 3 M Ammonium Sulfate solution (final concentration of ammoniumsulfate was 1M) to achieve a Bead: Protein ratio of 5 mg:100 μg. Thesolution was mixed gently and placed in a tube roller and incubated at37° C. for 16 hours.

Quantification of Coupling.

The tubes were taken out and subjected to a quick spin. The tubes wereplaced in the magna spin and the supernatant (60 μL) was collected(Supernatant). The beads were re-suspended in 60 μL of PBS and vortexedfor 10 seconds. The beads were allowed to settle in the magna spin andthe supernatant was collected as Wash 1 (W1) to remove the loosely boundproteins. The beads were washed three more times with 30 of PBS eachtime, and the supernatant was collected as W2, W3 and W4. The beads werefinally re-suspended in 25 μL of PBS and stored at 4° C. until use. Theamount of protein bound to beads was quantified by measuring the sum ofprotein left in the Supernatant, W1, W2, W3 and W4 through the Micro BCAmethod.

Micro BCA Method.

BSA was used as standard. The concentration range of BSA was 1 μg/mL to20 μg/mL in PBS. 10 μL of the Supernatant was mixed with 40 μL of PBS ina 96 well plate, and then with 100 μL of Micro BCA reagent. The platewas shaked at 37° C. for 3 hours. After incubation, the OD at 562 nm wasmeasured to determine the amount of protein in the Supernatant. Theamounts of protein in W1, W2, W3 and W4 were determined with the sameprocedure.

The amounts of protein in the Supernatant, W1, W2, W3 and W4 were addedand subtracted from the initial amount of the protein that was used forbeads coupling, which provided the amount of protein bound to the beadsand molarity of the protein was calculated.

αVβ6/αVβ8—LAP-TGF β1 Interaction:

αVβ6/αVβ8 coupled beads were treated with the ligand LAP TGF-β1 (LAP1)at room temperature for 3 hours. The complex (Integrin+Ligand) was thentreated with primary Ab (Anti-LAP1 Ab) overnight at 4° C. The wholecomplex (Integrin+Ligand+Primary Ab) was treated with Secondary Abconjugated with FITC and incubated for 2 hours. The complex was analyzedby either plate reader or Flow Cytometer.

10 μL of αVβ6/αVβ8 coupled beads were taken for the experiment. Theconcentration of integrins was 10 nM. 10 μL of LAP1 was taken (10 nM foraVβ6 and 20 nM for aVβ8). Reaction between integrin coupled beads andLAP1 was considered as the full reaction, and reaction without LAP1 or acompound of the disclosure was considered as the blank reaction. Thesamples were incubated in low protein binding tubes at room temperaturefor 3 hours. The tubes were briefly spun and placed in a Magna spin. Thesupernatant was removed. The beads were washed with assay buffer twiceto remove excess LAP1 and then re-suspended in 150 of assay buffercontaining 1:200 anti-LAP1 Ab (primary Ab). The tubes were placed in atube roller and incubated at 4° C. overnight. After a brief spin, thetubes were then placed in a Magna spin and the supernatant was removed.The beads were washed with assay buffer twice to remove excess primaryAb and then re-suspended in 150 μL of assay buffer containing 1:500secondary Ab conjugated with FITC. The tubes were incubated at roomtemperature for 2 hours in a tube roller. After a brief spin, the tubeswere placed in a Magna spin and the supernatant was removed. The beadswere washed with assay buffer twice followed by PBS. The beads were thenre-suspended in 300 μL of PBS and analyzed by a Flow Cytometer (BDFACSCalibur, Software—BDcell Quest Pro Version 6).

αVβ3/αVβ5—LAP-TGF β1 Interaction:

αVβ3/αVβ5 coupled beads were treated with the ligand at room temperaturefor 3 hours. The complex (Integrin+Ligand) was then treated withAnti-Vitronectin Ab conjugated with FITC overnight at 4° C. The complexwas analyzed by either plate reader or Flow Cytometer.

10 μL of αVβ3/αVβ5 coupled beads were taken for the experiment. Theconcentration of integrins was 10 nM. 10 μL of vitronectin was taken.The concentration was 10 nM. Reaction between integrin coupled beads andvitronectin was considered as the full reaction, and reaction withoutvitronectin or a compound of the disclosure was considered as the blankreaction. The samples were incubated in low protein binding tubes atroom temperature for 3 hours. The tubes were briefly spun and placed ina Magna spin. The supernatant was then removed. The beads were washedwith assay buffer twice to remove excess vitronectin and thenre-suspended in 150 μL of assay buffer containing 1:500 Anti-vitronectinAb conjugated with FITC. The tubes were placed in a tube roller andincubated at 4° C. overnight. After a brief spin, the tubes were placedin a Magna spin and the supernatant was discarded. The beads were washedwith assay buffer twice followed by PBS. The beads were thenre-suspended in 300 of PBS and analyzed by a Flow Cytometer (BDFACSCalibur, Software—BD Cell Quest Pro Version 6).

Quantification:

The samples were acquired using a BD FACSCalibur system and analyzedwith BD Cell quest pro Version 6. Median values for the following wereextracted from the software: Full reaction (Integrin+Ligand) with orwithout compound, Control: Without Ligand (LAP1/Vitronectin), andVehicle Control: Full reaction with DMSO. Blank=Test Medianvalue−control Median value. Percentage Inhibition=100−[(Blanked TestMedian/Blanked vehicle Median)*100]. Percentage of binding wascalculated with respect to full reaction. The value was subtracted from100 to get percentage of inhibition. All the plotted values were averageof triplicates. SD was determined for each experiment. IC₅₀ wasdetermined with Graph Pad Prism.

Inhibition of Integrin-Ligand Interaction by Reference Inhibitors:

The optimized protocol was validated by employing reference compoundssuch as Cilengitide (αVβ3/αVβ5-VN interaction) and CWHM12(αVβ6/αVβ8-LAP1 interaction). The full reaction (Integrin-LigandInteraction) was optimized as above. Integrin coupled beads were takenfor the experiment.

2 μL of 10 nM/20 nM of Ligand was taken and mixed with 8 μL of thecompound (i.e., Cilengitide or CWHM12, each diluted from a 10 mM stock).Reaction, with or without DMSO (0.08%), between Integrin and Ligand inthe absence of the compound was considered as the full reaction.Reaction with DMSO (0.08%) in the absence of compound and Ligand wasconsidered as the blank reaction.

The samples incubated in low protein binding tubes at room temperaturefor 3 hours. The tubes were placed in a Magna spin and the supernant wasdiscarded. The beads were washed with assay buffer twice to remove theexcess Ligand and then re-suspended in 150 μL of assay buffer containingthe primary antibody (1:500 of Anti-VN-FITC or 1:200 of Anti-LAP1 Ab).The tubes were placed in a tube roller and incubated at 4° C. overnight.After a brief spin, the tubes were placed in a Magna spin, and thesupernatant was discarded. In the case of αVβ3/αVβ5-VN interaction, thebeads were washed with assay buffer twice and finally washed with PBS.The beads were then re-suspended in 300 μL of PBS and analyzed by a FlowCytometer. In the case of αVβ6/αVβ8-LAP1 interaction, the beads werewashed with assay buffer twice and treated with 150 μL of SecondaryAntibody (1:500) for two hours at room temperature, washed twice withassay buffer and PBS, and finally re-suspended in 300 μL of PBS andanalyzed by a Flow Cytometer.

Table 3 shows the integrin inhibition activity of compounds of theinvention.

TABLE 3 Integrin Inhibition Assay Results αvβ6 αvβ8 αvβ8/ αvβ3 Cmpd #IC₅₀ (nM) IC₅₀ (nM) αvβ6 IC₅₀ (nM) A30s 9.57 17.56 1.83 A1 8.30 13.201.59 A4 5.69 21.35 3.75 1.4 A2 11.64 78.14 6.71 9.0 A3 18.16 79.31 4.370.57 A5 9.97 63.90 6.41 65.7 A6 8.25 24.02 2.91 190.1 A7 11.63 80.636.93 A23 14.80 150.70 10.18 4.3 A24 17.51 35.37 2.02 A28 16.88 34.392.04 A15s 9.28 5.92 0.64 A21 7.64 88.16 11.54 A21-1 NA NA NA A21-2 4.6050.30 10.93

Example 14. Anti-Angiogenic Activity Using Chick ChorioallantoicMembrane (CAM) Assay

CAM surfaces were grafted with gelatin sponges impregnated with theconcentrations of test compounds and 50 ng VEGF dissolved in PBS.Untreated CAM received only VEGF and PBS. Error bars represent SEM, N=5,P values for the treated groups were calculated by comparing with theuntreated group (*p<0.05, **p<0.01, ***p<0.001).

Test Substance Preparation:

Test samples and standards were dissolved in PBS and sterilized bypassing through a syringe filter (0.22 μm). hVEGF (SIGMA) 50 ng/μ1 wasprepared in sterile PBS.

Grafting:

Gelatin sponge (Abogel) was cut in approximately 2 mm³ pieces and loadedwith required test substance or PBS and VEGF. The graft was placed onthe CAM.

Eggs:

Fertile hen eggs were procured from a hatchery and were cleaned anddecontaminated using alcohol. 1 ml of albumin was removed using asyringe and incubated for 8 days. Grafts were placed on developing CAMsand further incubated to day 12. On day 12, CAMs were fixed with 4%formaldehyde in PBS, dissected and imaged.

Imaging:

Fixed CAMs were imaged under constant illumination and magnificationunder a stereomicroscope fitted with a digital camera (CANON).

Image Analysis:

Images were analyzed on MS PowerPoint keeping the image size constant. Aring was drawn around the graft and the size was kept constant. Bloodvessels crossing the ring were counted for each test group.

Statistical Analysis:

Data were analyzed on MS Excel 2007.

Example 15. Distribution in Plasma, Aqueous Humor, Vitreous Humor, andRetina after Topical Ocular Administration in Dutch Belted Rabbits

The plasma concentrations and ocular distribution (aqueous humor,vitreous humor, and retina) of Compounds A1, A2, and A3 were determinedfollowing topical ocular administration in Dutch Belted rabbits. Thetest compounds were administered in each eye at a volume of 50 μL/eye ata concentration of 1.0-2.5 mg/mL (Compound A2, 1.0 mg/mL; Compounds A1and A3 at 2.5 mg/mL). Plasma and different ocular tissue samples werecollected at pre-determined time points (1.0 and 8.0 hours for compoundA1; 0.5 and 8 hours for Compounds A2 and A3). Aqueous humor, vitreoushumor, and retina were collected from each eye at each time pointpost-dose. Also, weights were recorded. Plasma and ocular sampleconcentrations of the compounds were determined by LC-MS/MS.

Animal Dosing:

The exposure of Compounds A1, A2, and A3 was evaluated in Dutch Beltedrabbits. The study was not blinded. Each compound was dosed as n=3/timepoint for a total of nine rabbits. Rabbits were housed one per cage.Animals were not fasted, and food and water were supplied ad libitum.

Animals were anesthetized following the 13IA5 IACUC protocol for thedosing. Each rabbit received a bolus dose of test formulation viatopical ocular administration into both eyes at time zero on the day ofdosing. Plasma and ocular samples were collected at pre-determined timepoints. Animals for the 30-minute and 1-hour time points wereanesthetized for the entire duration of the study. The animals for the8-hour time point were recovered after dosing and then euthanized forsampling purposes.

At each time point, approximately 0.5 mL of blood was collected andplaced into chilled Na-heparin tubes containing citric acid. Bloodsamples were centrifuged at a speed of 3,000 g for 5 minutes to obtainplasma as quickly as possible. Samples were stored frozen at −80° C.until analysis. Animals were euthanized per the 13IA5 IACUC protocol andboth eyes were enucleated immediately. Following enucleation, each eyewas rinsed with PBS. Ocular samples from both eyes of each animal werecollected and weights were recorded. All the samples were frozenimmediately on dry ice, and stored at −60° C. to −80° C. for analysis.

Analysis of Plasma and Ocular Samples:

An LC-MS/MS method was developed for the determination of theconcentration of Compounds A1, A2, and A3 in rabbit plasma and ocularsamples. A pre-study standard curve was analyzed to determine thespecificity, range, and lower limit of quantitation of the method.

The following examples of ophthalmic formulations are given by way ofillustration:

Example 16. Evaluation of the Safety and Efficacy of Topically AppliedTest Compounds in the Laser-Induced Choroidal Neovascularization (CNV)Model in Dutch Belted Rabbits

Healthy male animals weighing between 1.5 kg and 2.0 kg were used inthese studies. Animals were weighed prior to dosing and at euthanasia,and more often if needed. Baseline fundus photography and fluoresceinangiography was performed on each animal prior to CNV induction.

Animals were anesthetized with an intramuscular injection of ketaminehydrochloride (20 mg/kg) and xylazine (2 mg/kg) for CNV induction,fundus photography, fluorescein angiography, and intravitreal (IVT)injections. Rabbits were maintained on isoflurane (approximately 1 to3%) in oxygen (approximately 1 to 2 L/min) as necessary. One drop oftopical proparacaine hydrochloride anesthetic (0.5%) was placed in eacheye before procedures. Additional topical ocular anesthesia was utilizedduring the procedure if needed.

CNV was induced by laser photocoagulation treatment. An external diodelaser was applied to the retina using a laser contact lens and a slitlamp biomicroscope. On Day 1, both eyes of each animal underwent laserphotocoagulation treatment using the following laser settings:

Number of Spots: 12-15 spots per eye

Power Range: 50-200 mW

Spot Size: 20-100 μm

Time: 0.05-0.1 seconds

Following laser treatment, 50 μL of a 25-μg/mL VEGF solution (1.25 μgdose) was intravitreally injected into each eye. Daily gross ocularexams were performed throughout the study period.

Clinical ophthalmic exams (slit-lamp biomicroscopy and indirectophthalmoscopy), fundus photography, and fluorescein angiography wereperformed at baseline and then weekly for up to 6 weeks post-induction.Exams were scored using the McDonald-Shadduck Score System. OpticalCoherence Tomography OCT imaging was performed weekly for diagnosticimaging during the exams.

On the last day of the study, blood sampling was performed just prior toadministration of the AM dose and at 2 hours post dosing. Blood sampleswere centrifuged at a speed of 3,000 g for 5 minutes to obtain plasma asquickly as possible. Samples were stored frozen at −80° C. untilanalysis. At the conclusion of the study, animals were euthanized perthe 13C232Q3 IACUC protocol and both eyes enucleated immediately.Following enucleation, each eye was rinsed with phosphate-bufferedsaline. Ocular samples (aqueous humor, vitreous humor retina andchoroid) from both eyes of each animal were collected and weights wererecorded. All the samples were frozen immediately on dry ice, and storedat −60° C. to −80° C. for analysis.

Example 17. Diagnosing Fibrosis

Fibrosis is a pathophysiological process in response to tissue injurydue to viral or bacterial infection, inflammation, autoimmune disease,trauma, drug toxicity, and so on. During this process, an excess amountof collagen is expressed and fibrous material forms in the extracellularspace of the affected tissue. Thus, fibrosis can be generally recognizedbased on the distinct morphology of fibrous tissue in a biopsy of theorgan in which fibrosis is suspected. Other means for detecting thepresence of fibrosis or developing fibrosis include computerized axialtomography (CAT or CT) scan, ultrasound, magnetic resonance imaging(MRI), and monitoring the level of one or more serum markers known to beindicative of fibrosis (e.g., various types of collagens).

The precise manner of diagnosing fibrosis also varies depending on theorgan where the fibrotic process takes place. For instance, biopsies aregenerally effective for diagnosing fibrosis of most organs, whereasendoscopy involving a fiber optic instrument (e.g., a sigmoidoscope or acolonoscope) can be a less traumatic alternative to detect fibrosis ofcertain organs such as the intestine.

Biopsy for Detecting Fibrosis

Standard procedures have been established for obtaining biopsy from agiven organ or tissue. For example, a specimen can be obtained duringexploratory surgery, but is more often obtained by inserting a biopsyneedle through the skin and into the organ or tissue. Before thisprocedure is performed, the person receives a local anesthetic.Ultrasound or CT scans may be used to locate the abnormal area fromwhich the specimen is to be taken.

Upon obtaining an organ or tissue biopsy, the sample is examined andgiven a score to indicate the presence and level of fibrosis in thesample. Most frequently used scoring systems include the METAVIR ormodified HAI (ISHAK) scoring system. The Knodell scoring system can alsobe used for analyzing the liver sample. The criteria used in scoring arewell established and known to those of skilled in the art. For example,the METAVIR system provides five gradings: F0 indicates the absence offibrosis; F1 indicates portal fibrosis without septa; F2 indicatesportal fibrosis and some septa; F3 indicates septal fibrosis withoutcirrhosis; and F4 indicates the presence of cirrhosis.

Biopsy is not only useful for the diagnosis of fibrosis, it can also aidphysicians to assess the effectiveness of fibrosis treatment/preventionmethods of the present invention by monitoring the progression offibrosis using methodologies known in the art. See, e.g., Poynard etal., Lancet 349:825, 1997.

Fibrosis Markers

There are numerous known serum markers whose level can be indicative ofthe presence and/or severity of fibrosis. Blood tests measuring markers,e.g., hyaluronic acid, laminin, undulin (type IV collagen) pro-peptidesfrom types I, II, and IV collagens, lysyl oxidase, prolyl hydroxylase,lysyl hydroxylase, PIIINP, PICP, collagen VI, tenascin, collagen XIV,laminin P1, TIMP-1, MMP-2, α2 macroglobulin, haptoglobin, gamma glutamyltranspeptidase, γ globulin, total bilirubin, apolipoprotein A1, etc.,according to the established methods can thus be useful for both thediagnosis of fibrosis and monitoring of fibrosis progression. Additionalmarkers, such as nucleic acid markers, can be used for detecting and/ormonitoring fibrosis. For instance, Wnt-4 has recently been indicated inlaboratory experiments as a gene that plays an important role in renalfibrosis, where its mRNA expression is significantly increased in thefibrotic tissue in the kidney (See, e.g., Surendran et al., Pediatr.140:119-24, 2002). The quantitative detection of gene expression of thistype of markers can be useful in the diagnosis and monitoring offibrosis

Example 18. Bleomycin Induced Mouse Pulmonary Fibrosis Model

Ninety-five male C57BL/6 mice were randomly and prospectively assignedto one group of fifteen animals and eight groups of ten animals each. Onday 0 and at least one hour prior to bleomycin induction, animals wereadministered the first dose of vehicle or test-article (i.e., a compoundof the present disclosure). At least one hour following dosing, all micewere anesthetized with isoflurane and placed on their backs lying on atable at approximately 60°. A small diameter cannula was inserted intothe trachea, and saline or bleomycin was slowly infused into the lungsin a volume of 40 μL.

Group 1 served as an untreated control group and received saline only(no bleomycin) on day 0. Groups 2-9 received 2.25 U/kg of bleomycin onday 0. The animals were then released into a recovery cage and allowedto wake up. From day 0 through day 21, treatments were administered onceor twice daily via oral gavage (PO). Vehicle treated animals (Group 2)received 0.4% methylcellulose. Remaining animals received eitherPirfenidone at 100 mg/kg (Group 3), Compound A15s at 100 mg/kg (Group4), 30 mg/kg (Group 5) or 10 mg/kg (Group 6), or Compound A21 at 100mg/kg (Group 7), 30 mg/kg (Group 8) or 10 mg/kg (Group 9).

All animals were weighed and evaluated daily for respiratory distress(defined as an increase in respiratory rate and/or obvious respiratoryeffort). Animals with severe respiratory distress, or animals that lostgreater than 30% of their total starting body weight, were euthanizedwithin 2 hours of observation.

On Day 21, prior to sacrifice, mice were anesthetized with IP injectionof ketamine/xylazine (100 mg/kg, 10 mg/kg). Once the animal wasdetermined to be non-responsive a shallow 2 cm vertical incision wasmade starting from 1 cm below the chin. The trachea was isolated and atransverse cut was made between tracheal rings approximately half-waythrough the trachea. A tracheotomy was performed by the insertion of an18 gauge polyethylene cannula through the incision secured with surgicalsuture to the trachea. Following cannulation, the adapter end of thecannula was attached to the flexiVent mechanical ventilator. The animalwas ventilated at 10 ml/kg tidal volume (V_(T)), 150 breaths per minuteand 3 cm H₂O positive end expiratory pressure (PEEP). Following a2-minute acclimation period, lung volume was standardized with 1,6-second deep inflation to a pressure of 30 cm H₂O followed by 2pressure-volume measurements up to 40 ml/kg. Each animal then underwenta measure of total respiratory impedance by applying a 3-secondpseudorandom frequency oscillation to the airway opening at 3, 6, 9 and12 cm H₂O PEEP. If at any time during this procedure the animal becameresponsive as demonstrated by a response to stimuli or spontaneousbreathing efforts the animal received a supplemental dose of 50 mg/kgketamine.

Compounds A15s and A21 reversed bleomycin induced lung stiffness at alldoses tested, as compared to the vehicle control group. The mid- andhigh-dose groups of Compound A15s produced a significant reversal ofbleomycin induced lung stiffness, and both groups were superior to thepositive control pirfenidone given at 100 mg/kg BID. The mid-dose groupof Compound A15s was indistinguishable from the saline treated animalgroup (i.e., animals not treated with bleomycin). The mid- and high-dosegroups of Compound A21 also produced similar, if not better, reversal ofbleomycin induced lung stiffness, as compared to the positive controlpirfenidone given at 100 mg/kg BID.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments and methods described herein. Such equivalents are intendedto be encompassed by the scope of the present invention.

All patents, patent applications, and literature references cited hereinare hereby expressly incorporated by reference.

The invention claimed is:
 1. A compound selected from the groupconsisting of:

or a pharmaceutically acceptable salt or solvate thereof.
 2. A compoundselected from the following or a pharmaceutically acceptable saltthereof:


3. The compound of claim 2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 4. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 5. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 6. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 7. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 9. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 10. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 11. The compound of claim2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 12. A pharmaceuticalcomposition comprising a compound of claim 2 and a pharmaceuticallyacceptable carrier or excipient.
 13. A pharmaceutical compositioncomprising a compound of claim 3 and a pharmaceutically acceptablecarrier or excipient.
 14. A pharmaceutical composition comprising acompound of claim 6 and a pharmaceutically acceptable carrier orexcipient.
 15. A pharmaceutical composition comprising a compound ofclaim 8 and a pharmaceutically acceptable carrier or excipient.
 16. Apharmaceutical composition comprising a compound of claim 10 and apharmaceutically acceptable carrier or excipient.
 17. A method oftreating fibrosis in a subject, comprising administering to a subject inneed thereof a therapeutically effective amount of a compound of claim 2to treat the fibrosis.
 18. A method of treating fibrosis in a subject,comprising administering to a subject in need thereof a therapeuticallyeffective amount of a compound of claim 3 to treat the fibrosis.
 19. Amethod of treating a disease or condition selected from maculardegeneration, age-related macular degeneration, diabetic retinopathy,diabetic macular edema, retinopathy of prematurity, or macular edemafollowing retinal vein occlusion in a subject, comprising administeringto a subject in need thereof a therapeutically effective amount of acompound of claim 2 to treat the disease or condition.
 20. A method oftreating a disease or condition selected from macular degeneration,age-related macular degeneration, diabetic retinopathy, diabetic macularedema, retinopathy of prematurity, or macular edema following retinalvein occlusion in a subject, comprising administering to a subject inneed thereof a therapeutically effective amount of a compound of claim 3to treat the disease or condition.